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Scientia Horticulturae 154 (2013) 102–108

Contents lists available at SciVerse ScienceDirect

Scientia Horticulturae

journa l h o me page: www.elsev ier .com/ locate /sc ihor t i

ffect of diurnal variability and storage conditions on essential oil content anduality of damask rose (Rosa damascena Mill.) flowers in north western Himalayas

akesh Kumar ∗, Saurabh Sharma, Swati Sood, Vijai K. Agnihotri, Bikram Singhatural Plant Products Division, CSIR-Institute of Himalayan Bioresource Technology (Council of Scientific and Industrial Research), Post Box No. 6, Palampur 176 061, HP, India

r t i c l e i n f o

rticle history:eceived 17 December 2011eceived in revised form 2 February 2013ccepted 4 February 2013

eywords:osa damascenaiurnal variabilitytorageose oil

a b s t r a c t

Field experiments were conducted at CSIR-Institute of Himalayan Bioresource Technology, Palampur,India to study the effect of diurnal variability and storage conditions of flower on oil content and com-position of damask rose (Rosa damascena Mill.) during 2011. In this study, the rose oil was obtained byhydrodistillation in Clevenger type apparatus and the components in the oil were analyzed by GC/MS. Theessential oil content and composition were affected by harvest time and storage conditions. The highestessential oil content (0.043%, v/w) was obtained from the rose flowers which were harvested at 04:00am and the lowest (0.017%, v/w) from the flowers harvested at 02:00 pm. The percentage of citronel-lol + nerol, main components of rose oil, increased with delay in harvesting. Geraniol content (26.3%) wasmaximum when the flowers were harvested at 10:00 am, but after that there was significant reductionin its concentration upto 06:00 pm. Storage duration of flowers at different temperature also affectedthe oil content and composition. There was 8.5% and 27.6% reduction in oil content when the flowerswere stored for 24 h at 4 ◦C, and 18 ± 1 ◦C or 25 ± 1 ◦C, respectively. Parallel to the increase of the storagetime citronellol + nerol content increased. The rates of hexadecane, nonadecane and methyl eugenol in

the flowers distilled immediately were determined to be lower than the stored flowers. The percentageof geraniol in the flowers distilled immediately were between 27.4% and declined to 4.4%, 6.9% and 18.1%after 24 h of storage at 25 ± 1 ◦C, 18 ± 1 ◦C and at 4 ◦C temperature, respectively. The optimal results interms of its oil content and components were obtained from the rose flowers distilled immediately afterthe harvest compared to the flowers stored at 25 ± 1 ◦C, 18 ± 1 ◦C. However, at 4 ◦C the flowers can bestored for 16 h without much effect in oil content and composition.

© 2013 Elsevier B.V. All rights reserved.

. Introduction

Damask rose (Rosa damascena Mill.) is the most importantpecies among the scented roses, yields a highly fragrant commer-ially valuable essential oil which is used in the perfumery industryGuterman et al., 2002). It is cultivated for its essential oil and

edicinal aspects in many areas of the world, e.g. Bulgaria, Turkey,ndia, and Iran (Tabaei-Aghdaei et al., 2006). The most importantroducts obtained from damask rose are rose oil, rose water, roseoncrete and rose absolute (Lawrence, 1991). R. damascena is gener-lly preferred for its highly prized rose oil, which is commonly usedn perfumery, cosmetics, beverages, soft drinks, ice-creams, and

s a fragrance component in ointments and lotions etc (Douglas,993). Besides its application in perfumery industry, researchersave also reported some valuable characteristics of damask rose

∗ Corresponding author. Tel.: +91 1894 233341; fax: +91 1894 230433.E-mail addresses: rakeshkumar@ihbt.res.in, rakeshkrana@yahoo.com

R. Kumar).

304-4238/$ – see front matter © 2013 Elsevier B.V. All rights reserved.ttp://dx.doi.org/10.1016/j.scienta.2013.02.002

oil such as anti-HIV, antimicrobial, antibacterial and antioxidantactivities (Mahmood et al., 1996; Ozkan et al., 2004; Basim andBasim, 2004). Damask rose oil soothes and harmonizes the mindand helps with depression, anger, grief, fear, nervous tension andstress. Flower yield is a complex trait, which is dependent on yieldcomponents and is highly influenced by many genetic as well asenvironmental factors (Yousefi et al., 2009).

Essential oils obtained from aromatic plants, are complex mix-tures of several chemical compounds including terpenes, alcohols,aldehydes and phenols. Rose oil, obtained from the flowers of R.damascena is rich in rose oxides, linalool, geraniol, citronellol andnerol (Moein et al., 2010). The main quality constituents of rose oilare citronellol and geraniol (Farooqi et al., 1988). Due to lack of nat-ural and synthetic substitutes and low oil content, rose oil is oneof the most expensive essential oil in the world markets. The mainfactors to be accounted on harvesting aromatic plants are the har-

vesting time, drying temperature, and period of drying (Jose et al.,2006). Awareness on factors that influences the yield and contentof essential oil is important especially for the producers (Ames andMathews, 1968).

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The chemical evaluation of damask rose oil has been done byany researchers (Sood and Singh, 1992; Moein et al., 2010). But

o one has studied the effect of diurnal variability on essential oilontent of damask rose. Yilmaz et al. (2011) have studied the effectf harvesting hour on some physical and mechanical properties ofamask rose. Delay in harvesting or transport to distillery results

n decreasing essential oil and quality. Mohamadi et al. (2011) havetudied the effect of different storage conditions on oil content andomposition of R. damascena petals.

Damask rose flowers annually once in hilly areas and its flower-ng period lasts for about 35–45 days (from the first week of Aprilo second half of May) in northern India. During peak season flow-rs collected at large quantities in the morning and there is overfillr when some technical fault occurs in the processing unit, flow-rs are not distilled immediately, fermentation starts in the flowersue to excessive temperature until distillation (Baser, 1992). Therere considerable losses of essential oil yield and quality in fer-ented petals (Baydar and Gokturk Baydar, 2005). Harvesting and

ost-harvesting steps are important for herbal and aromatic plantso obtain higher essential oil content and better quality. Time ofarvesting and processing are important non monetary agronomic

nputs which influences the essential oil yield and composition. Inhe literature there is limited data available for the role of diurnalield and chemical composition of essential oil (Angelopoulou et al.,002; Yaldiz et al., 2005; Gurbuz et al., 2005; Gumuscu et al., 2008).hus the objective of the study was to evaluate the effect of diurnalariability and storage conditions on oil content and compositionf damask rose flowers.

. Materials and methods

.1. Local climate

The present study was conducted at the experimental farmf CSIR-Institute of Himalayan Bioresource Technology (Councilf Scientific and Industrial Research), Palampur (1325 m amsl,2◦06′05′′N, 76◦34′10′′E), India during 2011. The site experiences

mean annual temperature of 18 ◦C. The soil of the experimentalite was clayey in texture, acidic in reaction (pH 6.1), high in organicarbon (1.05%), low in available N (150.5 kg ha−1), high in available

(21.2 kg ha−1) and available K (534.2 kg ha−1). During floweringeriod of damask rose maximum temperature ranges from 24.5 to4.5 ◦C, minimum temperature ranges from 15.5 to 22.5 ◦C, relativeumidity varies between 59% and 91%, evaporation from 3 to 9 mmnd sunshine hours ranges from 4 to 11.5 h.

.2. Experimental details

The rose plants used for these studies were transplanted inugust 2008 at a spacing of 1.5 m × 0.75 m. For the first experi-ent, the flowers were harvested at different time intervals viz.,

4:00 am, 06:00 am, 08:00 am, 10:00 am, 12:00 pm, 14:00 pm,6:00 pm and 18:00 pm to study the effect of diurnal variabilityn oil content and composition. In the second experiment, flow-rs were harvested at 06:00 am during May 2011. The flowerssed as control (unstored) were immediately subjected to distil-

ation, whereas, the other flowers were stored for 4, 8, 12, 16 20,4 h at different temperature conditions viz., 4 ◦C, 18 ± 1 ◦C and5 ± 1 ◦C at 90 ± 5% relative humidity to detect the influence oftorage conditions on the essential oil content and composition.n each treatment 1 kg flowers were tightly packed in plastic bags

nd kept at 4 ◦C in cold room. Three replications were maintained.ach package weighing 1.0 kg flowers was regarded as replication.hese petals were hydro distilled in the Clevenger apparatus at 0,, 8, 12, 16 20, 24 h after storage. Similarly flowers were kept at

turae 154 (2013) 102–108 103

18 ± 1 ◦C and 25 ± 1 ◦C in cold room and stacked in bamboo bas-ket covered with wet muslin cloth and 1 kg flowers were hydrodistilled in the Clevenger apparatus at 0, 4, 8, 12, 16 20, 24 h afterstorage.

2.3. Essential oil extraction

In both experiments oil content was determined by distillingthree samples per treatment in Clevenger’s apparatus. One kilo-gram of rose flowers was placed in a distillation apparatus with 3 lof water and hydrodistilled for 5 h. The flower weight was taken atthe time of measurement. The oil content was measured as percent-age (v/w). The obtained rose essential oil was dried with anhydroussodium sulphate and then stored at 4 ◦C until analyzed by gas chro-matography (GC) and GC–MS analyses.

2.4. GC analysis and quantification

Gas chromatographic analyses were carried out on a ShimadzuGC-2010 gas chromatograph fitted with FID detector and a DB-5fused silica capillary column (30 m × 0.25 mm i.d.; 0.25 �m filmthickness). The operating condition were as follows: carrier gasnitrogen, with a flow rate of 2 ml min−1, the oven temperaturewas programmed as follows: 70 ◦C (4 min) and then 70–220 ◦Cat 4 ◦C min−1, injector and detector temperatures were set at240 ◦C.

The quantitative composition was obtained by peak area nor-malization and the response factor for each component wasconsidered to equal 1.

2.5. Gas chromatography–mass spectrometry

GC–MS analyses were carried out by GC–MS (QP2010 Shimadzu,Tokyo, Japan) equipped with AOC-5000 Auto injector and DB-5(SGE International, Ringwood, Australia) fused silica capillary col-umn (30 m × 0.25 mm i.d.; 0.25 �m film thickness). Temperaturewas programmed from 70 ◦C for 4 min and then 220 at 4 ◦C min−1

and held for 5 min, “Injector temperature, 240 ◦C; Interface tem-perature, 250 ◦C”, acquisition mass range, 800–50 amu; ionizationenergy, 70 eV. Helium was used as a carrier gas with 1.1 ml min−1

flow rate.

2.6. Identification of components

The retention indices were calculated for all volatile con-stituents using homologous series of n-alkanes (C8–C24). Thecomponents of oil were identified by comparing their mass frag-mentation pattern with those stored in the computer librarynamely Wiley, New York mass spectral (MS) library, National Insti-tute of Standards and Technology, NIST (Stein, 2005), and theirretention indices (RI).

2.7. Statistical analysis

The first experiment was set up according to Randomized BlockDesign with 3 replications and each package weighing 1.0 kg flow-ers was regarded as replication at each harvesting. The secondexperiment was also laid out in a Randomized Block Design with3 storage conditions and 7 storing hours with three replications.Statistical analysis of data was done as per the standard analysis

of variance technique for the design used, and treatment meanswere compared at P = 0.05 level of significance. The data was ana-lysed by software SYSTAT-12 (SYSTAT Software Inc., Chicago, IL,USA).

104 R. Kumar et al. / Scientia Horticulturae 154 (2013) 102–108

) of R

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Fig. 1. Effect of harvesting time on oil content (%

. Results and discussion

.1. Effect of diurnal variability

.1.1. Oil yieldThe essential oil content of rose flowers harvested at different

ime intervals ranged from 0.017% to 0.043% (Fig. 1). The highest

il content was found when the flowers were harvested between4:00 and 06:00 am (0.043%) after that there was reduction of2%, 21%, 21%, 60% in oil content when the flowers were harvested

able 1ffect of harvesting time of flowers on oil composition of R. damascena (data from 1st exp

Sr. no. Harvesting time

Compounds RI 04:00 am 06:00 am 08:00

Oil (%, v/w) 0.043 0.043 0.0381 �-Pinene 932 1.4 0.5 0.5

2 �-Pinene 974 0.3 0.2 0.1

3 �-Myrcene 988 0.9 0.8 0.7

4 Linalool 1095 2.3 1.5 0.9

5 Phenyl ethyl alcohol 1110 2.6 1.4 1.1

6 Z-Rose oxide 1110 0.2 0.2 0.1

7 E-Rose oxide 1125 0.1 0.1 0.1

8 �-Terpineol 1180 0.3 0.3 0.2

9 �-Terpineol 1186 0.4 0.3 0.2

10 �-Citronellol +nerol 1229 31.5 28.2 23.7

11 Z-Citral 1238 0.4 0.3 0.3

12 E-Geraniol 1256 23.0 24.8 24.6

13 E-Citral 1268 0.2 0.2 0.2

14 Citronellyl acetate 1352 0.4 0.4 0.3

15 Eugenol 1356 2.7 1.7 1.5

16 Neryl acetate 1359 Tr 0.1 0.2

17 Geranyl acetate 1380 1.2 1.8 2.4

18 �-Bourbonene 1387 0.1 0.1 0.1

19 Methyl eugenol 1400 1.1 0.8 0.9

20 E-Caryophyllene 1417 0.6 0.7 0.6

21 �-Guaiene 1437 0.4 0.5 0.4

22 �-Humulene 1452 0.5 0.5 0.5

23 Germacrene D 1484 1.1 1.7 1.5

24 Pentadecane 1500 0.5 0.5 0.4

25 E,E-� Farnesene 1506 0.4 0.6 0.5

26 Hexadecane 1600 0.1 0.1 0.1

27 Heptadecane 1700 1.6 1.8 1.8

28 E,E-Farnesol 1720 0.4 0.9 1.1

29 Octadecane 1800 0.2 0.2 0.2

30 Nonadecane 1904 11.6 13.5 17.4

31 Eicosane 2000 1.0 1.1 1.5

32 Heneicosane 2112 1.8 5.4 7.6

Total 89.3 91.2 91.7

I: retention index; Tr: traces.

. damascena flowers (data from 1st experiment).

at 08:00 am, 10:00 am, 12:00 pm and 02:00 pm, respectively.The maximum reduction in oil content was at 02:00 pm whenthe temperature is maximum during the day after that constantincrease in oil content was observed at 04:00 pm (0.034%) and06:00 pm (0.034%). The reduction in oil content of flowers is dueto the increase in temperature, thereby evaporation of oil contentand reduction in weight of flowers. During the day of experiment

temperature ranges from 19.0 ◦C to 28.7 ◦C, and relative humiditydecreased from 67% to 48% from morning to evening. Baydar andGokturk Baydar (2005) and Yilmaz et al. (2011) also reported

eriment).

am 10.00 am 12:00 pm 02:00 pm 04:00 pm 06:00 pm

0.034 0.034 0.017 0.034 0.0341.2 0.3 0.6 Tr Tr0.3 0.1 Tr Tr Tr1.0 0.5 0.8 Tr 0.31.7 0.9 1.3 1.5 1.61.0 0.8 0.5 0.3 0.60.2 0.1 Tr Tr Tr0.1 0.1 Tr Tr Tr0.4 0.2 0.4 0.7 Tr0.4 0.2 0.3 0.8 0.4

22.4 17.1 19.2 20.2 23.40.3 0.3 Tr Tr Tr

26.3 18.2 11.2 14.8 9.30.2 0.2 0.3 Tr 0.30.4 0.4 0.4 0.2 0.52.1 1.6 1.1 2.2 1.30.3 Tr Tr Tr Tr3.0 2.4 1.4 0.7 1.00.1 0.2 Tr Tr Tr1.2 1.1 1.5 0.7 1.70.9 1.0 1.3 0.3 1.00.6 0.6 0.8 0.2 0.60.7 0.8 1.1 0.2 0.82.0 2.6 2.5 0.8 2.30.4 0.4 0.6 0.3 0.60.7 0.8 1.1 0.3 0.70.1 0.1 Tr 0.2 Tr1.2 1.7 1.3 2.7 2.31.2 1.6 0.6 1.3 0.60.2 0.2 Tr 0.3 Tr

13.1 20.5 13.2 26.5 23.81.2 2.1 1.5 2.3 2.56.4 7.6 12.5 11.7 16.0

91.3 84.7 75.5 89.2 91.6

R. Kumar et al. / Scientia Horticulturae 154 (2013) 102–108 105

Table 2Effect of different storage conditions on essential oil content (%) of R. damascena flowers (data from 2nd experiment).

Treatment Storage duration (h)

0 4 8 12 16 20 24 Mean

Storage temperature4 ◦C 0.047 0.047 0.047 0.047 0.047 0.043 0.043 0.04618 ± 1 ◦C 0.047 0.047 0.047 0.047 0.043 0.043 0.034 0.04425 ± 1 ◦C 0.047 0.047 0.047 0.047 0.047 0.034 0.034 0.043Mean 0.047 0.047 0.047 0.047 0.046 0.040 0.037LSD (P < 5%) for comparison of storage duration 0.0019

0.000.00

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R

LSD (P < 5%) for comparison of storage temperature

LSD (P < 5%) for comparison of interaction

SD: least significant difference.

imilar results under Turkish conditions. Earlier Weiss (1997)eported that the increase in the temperature cause removal of thessential oils from the trichomes of the petals.

.1.2. Oil compositionA comparative study of volatile components of R. damascena

ower oil harvested at different time intervals of the day is pre-ented in Table 1. The phenyl ethyl alcohol content, was higheruring early harvesting of flowers and decreased thereafter. High-st content of citronellol + nerol (31.5%) was observed when theowers were harvested at 04:00 am. Geraniol content increasedpto 10:00 am and decreased thereafter upto 02:00 pm. Rose oxidesere found upto 12:00 pm and diminished after that. Nonadecane,

icosane and heneicosane are not desirable constituents of rose oil

nd their percentage increased with delay in harvesting time ofose flowers. There are few studies on the effect of diurnal variabil-ty on essential oil of damask rose. High quantity of monoterpenelcohols (citronellol + nerol, geraniol, linalool, phenyl ethyl alcohol

able 3ffect of storage conditions on oil composition of R. damascena at 25 ± 1 ◦C (data from 2nd

Sr. no. Compounds RI 0 h 4 h

Oil (%, v/w) 0.047 0.047

1 �-Pinene 932 1.6 Tr

2 �-Pinene 974 0.5 Tr

3 �-Myrcene 988 2.2 0.7

4 Linalool 1095 1.8 0.8

5 Phenyl ethyl alcohol 1110 1.8 1.8

6 Z-Rose oxide 1110 0.1 Tr

7 E-Rose oxide 1125 0.1 0.2

8 �-Terpineol 1180 0.4 Tr

9 �- Terpineol 1186 0.4 0.2

10 �-Citronellol +nerol 1229 26.4 24.9

11 Z-Citral 1238 0.3 0.4

12 E-Geraniol 1256 27.4 20.0

13 E-Citral 1268 0.3 0.6

14 Citronellyl acetate 1352 0.4 0.5

15 Eugenol 1356 1.5 1.4

16 Neryl acetate 1359 0.5 0.3

17 Geranyl acetate 1380 2.3 2.0

18 �-Bourbonene 1387 Tr 0.1

19 Methyl eugenol 1400 0.7 1.1

20 E-Caryophyllene 1417 0.6 0.4

21 �-Guaiene 1437 0.3 0.2

22 �-Humulene 1452 0.4 0.3

23 Germacrene D 1484 1.1 1.0

24 Pentadecane 1500 0.4 0.4

25 E,E-� Farnesene 1506 0.4 0.5

26 Hexadecane 1600 Tr Tr

27 Heptadecane 1700 1.2 2.5

28 E,E-Farnesol 1720 1.0 1.2

29 Octadecane 1800 0.1 0.3

30 Nonadecane 1904 9.4 17.6

31 Eicosane 2000 0.8 1.6

32 Heneicosane 2112 3.8 9.0

Total 88.2 90.0

I: retention index; Tr: traces.

3052

and citral), which cause typical rosaceous and freshness character,and low quantity of hydrocarbons (nonadecane, eicosane and hene-icosane), which cause the solidification of oil, are desired to get highrose oil quality (Baser, 1992). In the present study monoterpenesdecreased with the delay of harvesting while reverse trend wasobserved for hydrocarbons. Among these monoterpene alcohols,citronellol + nerol and geraniol ranged from 17.1% to 31.5%, 9.3% to26.3%, respectively. Baydar et al. (2008) have also reported higherconcentrations of hydrocarbons in damask rose flowers stored forvarious duration under room conditions than the petals distilledimmediately.

3.2. Storage conditions

3.2.1. Oil contentThe results of this study showed that the oil content of the flow-

ers distilled immediately after harvest was higher (0.047%) thanstored flowers at all temperatures (Table 2). Storage temperature

experiment).

8 h 12 h 16 h 20 h 24 h

0.047 0.047 0.043 0.034 0.0340.2 Tr 0.5 0.3 0.7Tr Tr 0.1 0.2 0.20.7 0.3 0.4 0.2 0.31.1 0.4 0.7 0.9 0.62.1 1.2 2.0 1.5 1.70.2 0.2 0.4 0.5 0.80.2 0.2 0.4 0.4 0.40.1 0.1 0.1 Tr 0.10.3 0.1 0.2 0.2 0.235.0 33.6 39.0 41.1 39.90.4 0.2 0.2 Tr 0.215.9 8.2 7.3 4.0 4.40.6 0.2 0.2 0.2 0.10.7 0.7 0.6 0.6 0.61.1 0.8 0.6 0.5 0.80.2 0.1 Tr Tr 0.11.5 0.7 0.6 0.3 0.4Tr 0.1 Tr Tr 0.12.1 2.6 2.8 3.4 3.30.5 0.4 0.5 0.5 0.60.3 0.3 0.3 0.4 0.40.4 0.3 0.5 0.5 0.51.2 1.1 1.0 1.0 1.20.4 0.4 0.5 0.6 0.50.5 0.4 0.6 0.6 0.5Tr 0.1 Tr Tr Tr1.9 2.3 2.1 2.0 2.11.2 0.6 0.5 0.3 0.20.2 0.3 0.3 0.2 0.212.7 22.1 15.7 16.5 18.41.1 2.0 1.5 1.5 1.65.9 10.1 7.3 9.2 8.1

88.7 90.1 86.9 87.6 89.2

106 R. Kumar et al. / Scientia Horticulturae 154 (2013) 102–108

Table 4Effect of storage conditions on oil composition of R. damascena at 4 ◦C (data from 2nd experiment).

Sr. no. Compounds RI 4 h 8 h 12 h 16 h 20 h 24 h

Oil (%, v/w) 0.047 0.047 0.047 0.047 0.043 0.0431 �-Pinene 932 0.8 0.7 0.3 1.6 0.4 0.52 �-Pinene 974 0.2 0.2 0.2 0.3 0.1 0.13 �-Myrcene 988 1.3 0.9 1.3 1.4 0.8 0.84 Linalool 1095 1.3 0.9 1.0 1.5 0.8 0.95 Phenyl ethyl alcohol 1110 1.6 1.0 0.7 1.5 0.6 0.96 Z-Rose oxide 1110 0.3 0.2 0.2 0.4 0.3 0.37 E-Rose oxide 1125 0.2 0.1 0.4 0.3 0.1 0.28 �-Terpineol 1180 0.2 0.2 0.3 0.3 0.2 0.29 �-Terpineol 1186 0.3 0.2 0.3 0.3 0.2 0.2

10 �-Citronellol +nerol 1229 31.3 29.5 30.3 35.4 30.7 35.811 Z-Citral 1238 0.4 0.4 0.5 0.8 0.4 0.812 E-Geraniol 1256 24.6 21.0 17.6 20.4 15.3 18.113 E-Citral 1268 0.3 0.2 0.6 0.6 0.5 0.614 Citronellyl acetate 1352 0.5 0.4 0.3 0.4 0.4 0.415 Eugenol 1356 1.4 1.1 0.7 1.1 0.5 1.216 Neryl acetate 1359 0.3 0.2 0.2 0.2 0.1 0.217 Geranyl acetate 1380 1.2 0.9 1.0 0.8 0.7 0.618 �-Bourbonene 1387 Tr 0.1 0.1 0.1 0.1 0.119 Methyl eugenol 1400 0.6 0.5 0.7 0.6 0.5 0.720 E-Caryophyllene 1417 0.3 0.4 0.4 0.4 0.4 0.421 �-Guaiene 1437 0.3 0.3 0.3 0.3 0.3 0.322 �-Humulene 1452 0.4 0.4 0.4 0.4 0.4 0.423 Germacrene D 1484 0.9 1.2 1.3 1.1 1.2 1.224 Pentadecane 1500 0.3 0.5 0.5 0.4 0.5 0.525 E,E-� Farnesene 1506 0.3 0.4 0.5 0.4 0.4 0.426 Hexadecane 1600 0.1 0.1 0.1 0.1 0.1 0.127 Heptadecane 1700 1.9 2.6 2.7 2.0 3.3 2.628 E,E-Farnesol 1720 0.8 1.3 0.9 0.6 1.1 0.829 Octadecane 1800 0.1 0.2 0.2 0.2 0.3 0.330 Nonadecane 1904 11.2 15.5 14.7 11.1 18.7 15.031 Eicosane 2000 0.8 1.2 1.0 0.8 1.3 1.132 Heneicosane 2112 4.8 6.1 5.5 4.3 6.6 5.9

89.0 88.9 85.2 90.1 87.3 91.6

RI: retention index; Tr: traces.

Table 5Effect of storage conditions on oil composition of R. damascena at 18 ± 1 ◦C (data from 2nd experiment).

Sr. no. Compounds RI 4 h 8 h 12 h 16 h 20 h 24 h

Oil (%, v/w) 0.047 0.047 0.047 0.047 0.040 0.0341 �-Pinene 932 0.8 0.2 0.4 0.2 0.8 0.22 �-Pinene 974 Tr 0.1 0.1 0.1 0.2 0.13 �-Myrcene 988 1.3 0.7 0.7 0.4 0.5 0.34 Linalool 1095 1.5 0.7 0.8 0.5 0.6 0.55 Phenyl ethyl alcohol 1110 1.4 0.9 1.5 0.9 1.4 1.36 Z-Rose oxide 1110 0.3 0.2 0.4 0.4 0.6 0.67 E-Rose oxide 1125 0.2 0.1 0.2 0.3 0.4 0.38 �-Terpineol 1180 0.3 0.2 0.2 0.2 0.2 0.29 �-Terpineol 1186 0.3 0.2 0.2 0.1 0.1 0.2

10 �-Citronellol +nerol 1229 35.9 32.1 37.8 36.1 41.7 43.211 Z-Citral 1238 0.4 0.3 0.2 0.3 0.2 0.312 E-Geraniol 1256 26.9 19.3 12.1 8.9 7.2 6.913 E-Citral 1268 0.2 0.2 0.3 0.2 0.1 0.114 Citronellyl acetate 1352 0.5 0.4 0.6 0.5 0.6 0.515 Eugenol 1356 1.1 0.9 1.0 1.0 1.1 1.316 Neryl acetate 1359 0.2 0.2 0.1 Tr Tr Tr17 Geranyl acetate 1380 1.5 1.0 0.8 0.5 0.3 0.418 �-Bourbonene 1387 0.1 0.1 Tr 0.1 0.1 0.119 Methyl eugenol 1400 0.7 0.9 1.6 1.9 3.0 2.820 E-Caryophyllene 1417 0.4 0.3 0.4 0.4 0.5 0.421 �-Guaiene 1437 0.3 0.2 0.3 0.3 0.4 0.322 �-Humulene 1452 0.3 0.3 0.4 0.4 0.5 0.423 Germacrene D 1484 1.0 1.0 1.0 1.1 1.3 1.224 Pentadecane 1500 0.3 0.4 0.4 0.4 0.4 0.525 E,E-� Farnesene 1506 0.3 0.3 0.4 0.4 0.4 0.426 Hexadecane 1600 0.1 0.1 0.1 0.1 0.1 0.127 Heptadecane 1700 1.5 2.7 2.2 3.0 2.5 2.528 E,E-Farnesol 1720 0.8 0.9 0.5 0.4 0.3 0.329 Octadecane 1800 0.1 0.2 0.2 0.3 0.2 0.230 Nonadecane 1904 8.8 18.0 14.7 21.5 17.2 17.431 Eicosane 2000 0.6 1.3 1.0 1.6 1.3 1.332 Heneicosane 2112 3.2 6.7 6.0 8.8 6.8 7.2

91.3 91.1 86.6 91.3 91.0 91.5

RI: retention index; Tr: traces.

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R. Kumar et al. / Scientia Ho

ignificantly affected the oil content. Flowers stored at 4 ◦Cecorded significantly higher oil than those flowers which weretored at 18 ± 1 ◦C and 25 ± 1 ◦C. With the increase in storage dura-ion a regular decline was observed in oil contents. Interactionffect of storage temperature and storage duration of damask roseowers on oil content was also significant (Table 2). Essential oilontent of damask rose flowers stored at 4 ◦C and 25 ± 1 ◦C for6 h, and at 18 ± 1 ◦C for 12 h, respectively, remained at par, butfter that it declined significantly. Thus, the flowers can be storedt 4 ◦C for 16 h and upto 12 h at 18 ± 1 ◦C to 25 ± 1 ◦C by coveringith muslin cloth without any effect in oil content. After that oil

ontent decreased by 8.6% and 27.6% and 27.6% with increase intorage duration upto 24 h at 4 ◦C, 18 ± 1 ◦C and 25 ± 1 ◦C, respec-ively. Baydar and Gokturk Baydar (2005) have also reported thathe oil content of the rose flowers distilled immediately after thearvest was high and that oil content decreased significantly ashe waiting duration for distillation increased. For instance, it waseported that the oil content in the flowers distilled immediatelyfter the harvest was 0.035% and that it became 0.030% 12 h later,.027% 24 h later and 0.025% 36 h later (Baydar and Gokturk Baydar,005). Mohamadi et al. (2011) have reported decrease in the essen-ial oil content of damask rose petals immersed in distilled waternd stored at 7 ◦C after two days.

.2.2. Oil compositionA perusal of data presented in Tables 3–5 revealed that stor-

ge duration also influenced the composition of oil. The significanthanges were observed in the percentage of citronellol + nerol anderaniol. With the increase in storage duration citronellol + nerolontent increased at 4 ◦C, 18 ± 1 ◦C and 25 ± 1 ◦C, whereas, geran-ol content decreased as the storage duration increased. Methylugenol content increased in opposition to the content of geran-ol when the duration of storage increased from 4 to 24 h while theate of geraniol and neryl acetate decreased. These findings are ingreement to those of (Baydar et al., 2008). Kazaz et al. (2009, 2010)nd Mohamadi et al. (2011) have also reported that citronellol,ncreased in the stored petals in comparison to the petals distilledmmediately. It was found that nonadecane and eicosane amongydrocarbons and methyl eugenol in the ester group increased inhe stored flowers in comparison to the flowers distilled imme-iately. Similar results were reported by Baydar et al. (2008) andazaz et al. (2009). Methyl eugenol is a high value aroma chemi-al used in cosmetic products and flavouring agents, however, it isot desired above a certain concentration in the essential oils dueo side effects on human health (Harris, 2002). Citronellol/geraniolC/G) ratio has been used for evaluating the odor quality of roseil (Kovats, 1987). The best odor of rose oil is produced when theatio is between 1.25 and 1.30 (Baser, 1992). Our results showedhat citronellol + nerol/geraniol ratio was 0.96, in the petals dis-illed immediately and 9.07, 6.26 and 1.98 in the flowers storedt 25 ± 1 ◦C, 18 ± 1 ◦C and 4 ◦C for 24 h. Baydar et al. (2008) havebserved that an increase in the internal temperatures of flowersesulted in significant removal of volatile substances from spe-ial parenchyma cells in the petals. Mohamadi et al. (2011) haveeported that chemical composition of rose oils changes depend-ng on time of storage. They further reported that the percentagef monoterpene alcohols increased from 39.92% to 49.23% whereasinalool and geraniol decreased after 5 days of storage at −20 ◦C.

In this study, the optimal results in terms of essential oil compo-ents were detected in the oils obtained from the flowers distilled

mmediately after the harvest. Though oil content was not affectedpto 16 h of harvest but oil composition was affected with increase

n storage duration even at low temperature of 4 ◦C. In this study,he best results in terms of rose oil content and quality werebtained from the flowers distilled immediately. Nevertheless, theetals are prevented from being distilled immediately since the

turae 154 (2013) 102–108 107

flowering period of damask rose is short. To prevent the loss ofquality, flowers were stored in wooden basket and covered withmoistened muslin cloth.

4. Conclusion

It can be concluded that the harvesting hour of damask roseflowers had significant effect on the yield of essential oil andchemical composition of various compounds. Harvesting flowersbetween 04:00 am and 06:00 am is beneficial for oil content andquality. The processing time of damask rose also had an effect oncomposition of essential oil showing the presence of various com-pounds in different periods. In order to increase the effectivenessof essential oil production, one should have the knowledge aboutthe nature of the plant and the causes of losses that may lead toboth wastage and reduction in yield and quality of the product. Inorder to get higher essential oil content, distillation should be madeas quickly as possible after harvest. The storage of rose flowers notonly reduces the oil content, but also increases the citronellol+ nerolcontent in opposition to the content of geraniol. It is generally pre-ferred that the citronellol content should be over 35% in the roseoil trade. But the oils from non-fermented petals generally con-tain citronellol lower than 35%. Thus storage of flowers for shortperiod should be allowed in order to increase the citronellol + nerolcontent.

Acknowledgements

The authors are grateful to the Director, IHBT, Palampur for pro-viding necessary facility during the course of study. The authors arealso thankful to Mr. Sushil Kumar and Vijaylata Pathania, technicalassistants, for field management and chemical analysis. Authors arealso grateful to Council of Scientific and Industrial Research, NewDelhi. This is IHBT Publication No. 2275.

References

Ames, G.R., Mathews, W.S.A., 1968. The distillation of essential oils. Trop. Sci. 10,136–148.

Angelopoulou, D., Demetzos, C., Perdetzoglou, D., 2002. Diurnal and seasonal vari-ation of the essential oil labdanes and clerodanes from Cistus monspeliensis L.leaves. Biochem. Syst. Ecol. 30, 189–203.

Baser, K.H.C., 1992. Turkish rose oil. Perfum. Flavor. 17, 45–52.Basim, E., Basim, H., 2004. Evaluation of antibacterial activity of essential oil of Rosa

damascena on Erwinia amylovora. Phytoparasitica 32, 409–412.Baydar, H., Gokturk Baydar, N., 2005. The effects of harvest date, fermentation

duration and Tween 20 treatment on essential oil content and composition ofindustrial oil rose (Rosa damascena Mill.). Ind. Crops Prod. 21, 251–255.

Baydar, H., Schulz, H., Kruger, H., Erbas, S., Kineci, S., 2008. Influences of fermenta-tion time, hydro-distillation time and fractions on essential oil composition ofDamask Rose (Rosa damascena Mill.). J. Essent. Oil Bear. Plants 11, 224–232.

Douglas, M., 1993. Rose-Rosa damascena ‘Trigintipetala’. Crop Food Res., 1–5.Farooqi, A.H., Abad Sharma, S., 1988. Effect of growth retardants on flowering of Rosa

damascena Mill. In: Sinha, S.K., Sane, P.V., Bhargava, S.C., Agrawal, P.K. (Eds.),Proceeding of International Congress of Plant Physiology, vol. 2. Society of PlantPhysiology and Biochemistry, New Delhi, pp. 1369–1372.

Gumuscu, A., Ipek, A., Sarihan, E.O., Gurbuz, B., Kaya, M.D., Arslan, N., 2008. Effectsof diurnal and ontogenetic variability on essential oil composition of oregano(Origanum vulgare var. hirtum). Asian J. Chem. 20, 1290–1294.

Gurbuz, B., Ipek, A., Basalma, D., Sarihan, E.O., Sancak, C., Ozcan, S., 2005. Effects ofdiurnal variability on essential oil composition of sweet basil (Ocimum basilicumL.). Asian J. Chem. 18, 285–288.

Guterman, I., Shalit, M., Menda, N., Piestun, D., Yelin, M.D., Shalev, G., Bar, E., Davydov,O., Ovadis, M., Emanuel, M., Wang, J., Adam, Z., Pichersky, E., Lewinsohn, E., Zamir,D., Vainstein, A., Weiss, D., 2002. Rose scent: genomics approach to discoveringnovel floral fragrance-related genes. Plant Cell 14, 2325–2338.

Harris, B., 2002. Methyl eugeneol – the current bete noire of aromatherapy. Int. J.Aromather. 12, 193–201.

Jose, L.S., Filho, C., Arie, F.B., Pericles, B.A., Polyana, A.D.E., Alberto, S.M., Socrates,

C.H.C., Arrigoni, B.M.F., Silva, M.R., 2006. Influence of the harvesting time, tem-perature and drying period on basil (Ocimum basilicum L.) essential oil. Rev. Bras.Farmacogn. 16, 24–30.

Kazaz, S., Erbas, S., Baydar, H., Dilmacunal, T., Koyuncu, M.A., 2010. Cold storage ofoil rose (Rosa damascena Mill.) flowers. Sci. Hortic. 126, 284–290.

1 rticul

K

K

L

M

M

M

O

08 R. Kumar et al. / Scientia Ho

azaz, S., Erbas, S., Baydar, H., 2009. The effects of storage temperature and durationon essential oil content and composition oil rose (Rosa damascena Mill.). Turk. J.Field Crops 14 (2), 89–96.

ovats, E., 1987. Composition of essential oils. Part 7. Bulgarian oil of rose (Rosadamascena Mill.). J. Chromatogr. 406, 185–222.

awrence, B.M., 1991. Progress in essential oil: rose oil and extracts. Perfum. Flavor.16, 43–77.

ahmood, N., Piacente, S., Pizza, C., Burke, A., Khan, A.I., Hay, A.J., 1996. The anti-HIV activity and mechanisms of action of pure compounds isolated from Rosadamascena. Biochem. Biophys. Res. Commun. 229, 73–79.

oein, M., Karami, F., Tavallali, H., Ghasemi, Y., 2010. Composition of the essentialoil of Rosa damascena Mill. from South of Iran. Iran. J. Pharm. Sci. 6, 59–62.

ohamadi, M., Mostafavi, A., Shamspur, T., 2011. Effects of storage on essential oil

content and composition of Rosa damascena Mill. petals under different condi-tions. J. Essent. Oil Bear. Plants 14, 430–441.

zkan, G., Sagdic, O., Baydar, N.G., Baydar, H., 2004. Antioxidant and anti bac-terial activities of R. damascena flower extracts. Food Sci. Technol. Int. 10,277–281.

turae 154 (2013) 102–108

Sood, R.P., Singh, B., 1992. Constituents of rose oil from Kangra Valley H.P. (India). J.Essent. Oil Res. 4, 425–426.

Stein, S.E., 2005. Mass Spectral Database and Software, Version 3.02. National Insti-tute of Standards and Technology (NIST), Gaitherburg, MD, USA.

Tabaei-Aghdaei, S.R., Hosseini Monfared, H., Fahimi, H., Ebrahimzadeh, H., Jebelly,M., Naghavi, M.R., Babaei, A., 2006. Genetic variation analysis of different pop-ulation of Rosa damascena Mill. in NW, Iran using RAPD markers. Iran. J. Bot. 12,121–127.

Weiss, E.A., 1997. Essential Oil Crops. CAB International, New York, USA.Yaldiz, G., Sekeroglu, N., Ozguven, M., Kirpik, M., 2005. Seasonal and diurnal vari-

ability of essential oil and its components in Origanum onites L. grown in theecological conditions of Cukurova. Grasas Aceites 56, 254–258.

Yilmaz, D., Ekinci, K., Dilmacunal, T., Erbas, S., 2011. Effect of harvesting hour on

some physical and mechanical properties of Rosa damascena Mill. J. Sci. FoodAgric., http://dx.doi.org/10.1002/jsfa.4351.

Yousefi, B., Tabaei-Aghdaei, S.R., Darvish, F., Assareh, M.H., 2009. Flower yield per-formance and stability of various Rosa damascena Mill. landraces under differentecological conditions. Sci. Hortic. 121 (3), 333–339.