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Extraction Methods for Tuberose Oil and Their Chemical

Components

Article  in  Kasetsart Journal - Natural Science · January 2009

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Kasetsart J. (Nat. Sci.) 43 : 204 - 211 (2009)

Extraction Methods for Tuberose Oil and

Their Chemical Components

Prapassorn Rakthaworn1*, Uraiwan Dilokkunanant1,

Udomlak Sukkatta1, Srunya Vajrodaya2, Vichai Haruethaitanasan3,
Potechaman Pitpiangchan1 and Putthita Punjee 1

ABSTRACT

The objectives of the project were to compare essential oil extraction methods from the double-
flower variety of tuberose (Polianthes tuberosa L.). The flowers were extracted by cold or hot enfleurage,
or by solvent extraction with hexane or petroleum ether. The chemical composition of the tuberose
absolutes was analyzed by gas chromatography-mass spectrometry (GC-MS). The results showed that
percentage yields of tuberose oil from cold enfleurage, hot enfleurage, hexane and petroleum ether
extractions were 0.3137%, 6.5808%, 0.0279% and 0.0182%, respectively. The main chemical component
detected in both enfleurage absolutes was methyl benzoate, while benzyl benzoate and pentacosane
were found to be the main chemical components in hexane and petroleum ether absolutes, respectively.
Key words: Polianthes tuberosa L., essential oil, absolute, distillation, enfleurage, solvent extraction,
chemical component

INTRODUCTION various plant organelles and as plant protection

against herbivores and infection, as well as to
The tuberose, Polianthes tuberosa L., is attract pollinators (Dudareva and Negre, 2005).
a tuberous perennial plant with a waxy, luminous At present, the demand for volatile oil is
white flower in the family Agavaceae. Its flower expanding. Essential oils can be extracted using a
odor is very sweet, floral and honey-like and can variety of methods, such as hydro distillation and
help give emotional strength and center the mind. solvent extraction, although some are not
It is known to improve an individual’s capacity commonly used today, such as cold and hot
for emotional depth and can stimulate the right enfleurage extraction (Maliga, 2003).
side of the brain and bring serenity to the mind The objectives of this work were to use
and heart. It also contains anti-inflammatory and tuberose flowers to compare the scent, percentage
antispasmodic properties (Maliga, 2003). oil yield and chemical component of tuberose
Tuberose flowers have long been used absolutes obtained by cold or hot enfleurage
in perfumery as a source of essential oils and aroma extraction and by solvent extraction using hexane
compounds. These aromatics are synthesized in or petroleum ether.

1 Herb and Biodiverstiy Technology Unit, KAPI, Kasetsart University, Bangkok 10900, Thailand.
2 Department of Botany, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
3 Department of Product Development, Faculty of Agro-Industry, Kasetsart University, Bangkok 10900, Thailand.
* Corresponding author, e-mail: aappsr@ku.ac.th
 Kasetsart J. (Nat. Sci.) 43(5) 205

MATERIALS AND METHODS flowers were filtered out and replaced with new
flowers. The steps were repeated using 300, 400,
Flower preparations 500, 600, 700 and 800 g of flower in each
Tuberose panicles were cut-off and put treatment. The flower scent was extracted from
into a water tank to keep the flowers fresh. Freshly the pomade by the same method used for the
opened blossoms were collected every day, and enfleurage extraction (Gupta, 1952).
weighed and subjected to extraction. Solvent Extraction: Hexane and
petroleum ether were used to extract the scents
Extraction methods from tuberose flowers because they are strongly
Enfleurage: Cold and hot enfleurage non-polar solvents and are frequently used in
processes were used in the tuberose oil extraction. solvents to extract oils. Six different weights of
In cold enfleurage, palm wax was heated to 80°C 100,150, 200, 250, 300 and 350 g of flowers were
for 2 h and poured into rectangular glass trays, soaked in 1 L of each solvent for 1 h. After
with 100 ml/tray. After the palm oil had cooled removing the debris, the solvents were evaporated
down and turned to wax at room temperature, leaving the concrete behind. Tuberose absolute
tuberose flowers were put on the wax in each tray was extracted from each concrete sample using
and covered with another waxed tray. The flowers alcohol. Physical appearances, such as color, odor
were replaced with fresh flowers every 24 h. Six and other characteristics were observed for all
pairs of the trays with 1,000, 1,500, 2,000, 2,500, extracts. Yields of extracts from each method were
3,000 and 3,500 g of tuberose flowers were studied assessed, compared, and used to determine the
as shown in Table 1. The floral scents in the wax saturation point of fat, oil and solvent to absorb
(pomade) were extracted by ethanol and the the scent from the tuberose flowers.
ethanol was evaporated leaving the absolute de
enfleurage behind. In hot enfleurage, tuberose Chemical composition analysis
flowers were put into 400 ml palm oil, which was Absolutes in petroleum ether and in
warmed at 60°C. The flowers were warmed for hexane 100 ppm were analyzed for their main
30 min and cooled down at room temperature. chemical components by a Shimadzu QP5050A
After leaving overnight at about 8-10°C, the palm gas chromatography mass spectrometer.
oil was warmed up to 60°C again and the previous

Table 1 Tuberose flowers used in enfleurage extractions.

Extraction method Tuberose flower weight (g)
Treatment 1 Treatment 2 Treatment 3 Treatment 4 Treatment 5 Treatment 6
Cold enfleurage 1,000 1,500 2,000 2,500 3,000 3,500
(200 ml wax / treatment)
Hot enfleurage 300 400 500 600 700 800
(400 ml oil / treatment)

Table 2 Tuberose flowers used in solvent extractions.

Extraction method Tuberose flower weight (g)
Treatment 1 Treatment 2 Treatment 3 Treatment 4 Treatment 5 Treatment 6
Hexane extraction 100 150 200 250 300 350
Petroleum ether extraction 100 150 200 250 300 350
206 Kasetsart J. (Nat. Sci.) 43(5)

RESULTS AND DISCUSSION easily removed from the ethanol than the palm oil
(Pensuk et al., 2007), which may have caused the
Extractions difference in yields. However, the cold enfleurage
Enfleurage: Tuberose absolutes of both scent was more similar to that of a fresh tuberose
cold and hot processes were similar in appearance; flower than the hot enfleurage absolute, because
they were sticky, waxy orange-brown oils. The tuberose oil is extremely delicate and thus, heating
absolute from the cold extraction was lighter in the petals would destroy the most delicate
color than from the hot extraction. The saturation components (Handa, 2005). Cold enfleurage
point of palm stearin (cold enfleurage) and palm absolute gave the scent most strongly reminiscent
oil (hot enfleurage) for the absorption of essential of natural tuberose flowers and was safe for use;
oil from tuberose flowers was 2,500 g flower/200 it was appropriate for use in high-grade perfume
ml palm wax and 500 g flower/400 ml palm oil, materials, However, enfleurage had the
respectively and the yield varied from treatment disadvantage of being very labor-intensive and
to treatment (Figure 1) with average yields of expensive.
0.3137% and 6.5808%, respectively. However, it Solvent extraction: The concretes and
has been noted that the saturation point of fat absolutes obtained from hexane and petroleum
depends on the essential oil content in the flowers ether extractions appeared similar. Concretes were
and properties of the fat (Pensuk et al., 2007). a yellow solid with a tuberose odor and absolutes
The absolute yields from cold enfleurage were brownish yellow with a strong tuberose scent.
were less than those from hot enfleurage in all The scent of tuberose absolute extracted from
treatments. This was probably due to the heat used petroleum ether was closer to that of natural
in the extraction process, as the wax was able to flowers than from hexane. The optimum amount
extract a greater yield from tuberose petals than of 150 g flower/l for hexane and petroleum ether
in the cold process (Alchemy Works, 2007). extraction produced yields of 0.0279% and
Moreover, absolutes from both methods contained 0.0192%, respectively (Figure 2). Solvent
palm wax and palm oil, which was extracted by extraction was more cost-efficient than the
ethanol used in the process. The wax was more enfleurage process, and so was appropriate for

Figure 1 Yield of tuberose oil from cold (a) and hot (b) enfleurage extractions.
(a) Treatment 1 = 1000 g flowers/200 ml wax (b) Treatment 1 = 300 g flowers/400 ml oil
Treatment 2 = 1500 g flowers/200 ml wax Treatment 2 = 400 g flowers/400 ml oil
Treatment 3 = 2000 g flowers/200 ml wax Treatment 3 = 500 g flowers/400 ml oil
Treatment 4 = 2500 g flowers/200 ml wax Treatment 4 = 600 g flowers/400 ml oil
Treatment 5 = 3000 g flowers/200 ml wax Treatment 5 = 700 g flowers/400 ml oil
Treatment 6 = 3500 g flowers/200 ml wax Treatment 6 = 800 g flowers/400 ml oil
 Kasetsart J. (Nat. Sci.) 43(5) 207

Figure 2 Tuberose concretes and absolutes obtained from hexane (a) and petroleum ether (b) extraction.
Treatment 1 = 100 g flowers/1,000 ml solvent Treatment 4 = 250 g flowers/1,000 ml solvent
Treatment 2 = 150 g flowers/1,000 ml solvent Treatment 5 = 300 g flowers/1,000 ml solvent
Treatment 3 = 200 g flowers/1,000 ml solvent Treatment 6 = 350 g flowers/1,000 ml solvent

producing absolute on a pilot scale, but, this was niobe and tuberose. It possesses a pleasant smell,
not considered the best method for the extraction strongly reminiscent of the fruit and is mostly used
of essential oils, as the solvent was harmful and in perfumery (Reverchon and Poletto, 1996). The
could leave a residue behind that could cause results showed that temperature and the method
allergies and effect the immune system (Handa, of extraction affected not only the chemical
2005). composition of the absolutes, but also the
percentage yield.
Chemical composition Solvent extraction: Fourteen chemicals
Enfleurage: The chromatogram of the were detected in the tuberose hexane absolute.
cold enfleurage absolute detected 10 chemicals Benzyl benzoate (24.25%), pentacosane (19.23%)
(Figure 3 and Table 3), with the major components and 7-decen-5-olide (14.96%) were the major
being methyl benzoate (30.17%), benzyl benzoate chemical components (Figure 5 and Table 5).
(23.64%), 7-decen-5-olide (13.33%) and methyl There were 14 chemicals identified in the
salicylate (12.11%). petroleum ether absolute. The main components
Ten chemical constituents were also were pentacosane (29.44%), 7-decen-5-olide
detected in the hot enfleurage absolute, with (18.13%) and heptacosane (12.53%). However,
methyl benzoate being the main component again, benzyl benzoate represented only 10.28% (Figure
but with a higher percentage yield (44.85%) than 6 and Table 6). Petroleum ether extracted more
in the cold enfleurage absolute. Moreover, (Z)-3- wax, e.g. pentacosane and heptacosane, from plant
hexenyl 2-oxopropanoate was a major component cells than hexane. Plant wax was located in the
(27.38%) only in the hot enfleurage absolute. cuticle of the epidermis. Some flowers such as
There were some differences in the chemical jasmine and tuberose contained scent in the flower
composition between the cold and hot enfleurage wax, which could be dissolved with some solvents
absolutes. However, methyl benzoate, methyl (Alchemy-works, 2007). If the wax content were
salicylate and benzyl benzoate were found in both high, it would be difficult to obtain the essential
absolutes. Among these components, methyl oil by distillation. In this case, extraction by
benzoate was considered the major characteristic solvents extracted more oil resulting in the
of tuberose absolute, which is known as oil of tuberose scent of the petroleum ether absolutes
208 Kasetsart J. (Nat. Sci.) 43(5)

Figure 3 Chromatogram of cold enfleurage absolute.

Table 3 Chemical compositions of cold enfleurage absolute.

Peak No. Retention time (min) % Relative peak area Possible compounds
1 10.996 30.17 methyl benzoate
2 12.896 12.11 methyl salicylate
3 14.567 1.78 indole
4 14.818 1.47 (E)-citral
5 15.297 4.45 methyl anthranilate
6 16.021 1.80 methyl eugenol
7 17.306 8.83 (E)-methyl isoeugenol
8 17.373 13.33 7-decen-5-olide
9 20.755 2.43 (z)-beta-farnesene
10 21.943 23.64 benzyl benzoate

Table 4 Chemical compositions of hot enfleurage absolute.

Peak No. Retention time (min) % Relative peak area Possible compounds
1 10.987 44.85 methyl benzoate
2 12.904 7.18 methyl salicylate
3 14.740 2.91 2,4-decadien-1-al
4 14.815 27.38 (Z)-3-hexenyl 2-oxopropanoate
5 15.736 4.15 1-tetradecene
6 17.300 1.78 (E)-methyl isoeugenol
7 17.374 3.35 (Z)-nerolidol
8 18.460 3.39 1-hexadecene
9 21.922 3.75 benzyl benzoate
10 23.119 1.26 2-heptadecanone

Figure 4 Chromatogram of hot enfleurage absolute.

 Kasetsart J. (Nat. Sci.) 43(5) 209

Figure 5 Chromatogram of tuberose hexane absolute.

Table 5 Chemical compositions of tuberose absolutes obtained by hexane extraction.

Peak No. Retention time (min) %Relative peak area Possible compound
1 14.804 3.70 (Z)-3-hexenyl 2-oxopropanoate
2 16.009 1.07 methyl eugenol
3 16.744 2.32 methyl isoeugenol
4 17.294 1.89 (E)-methyl isoeugenol
5 17.364 14.96 7-decen-5-olide
6 17.494 8.36 2,4,-di-tert-butylphenol
7 18.449 8.85 1-hexadecene
8 20.736 1.94 alpha-farnesol
9 21.901 24.25 benzyl benzoate
10 24.198 1.39 benzyl salicylate
11 26.245 4.96 ecosanol
12 32.440 2.47 tricosane
13 36.972 19.23 pentacosane
14 43.542 4.60 heptacosane

being closer to natural flowers than from hexane oilde, (Z)–6–dodecadien–4–oilde, eugenol,
absolute. In the current study, most of the farnesol, geraniol, hecogenin, methyl benzoate,
chemicals identified in from both solvent methylvanillin, nerol, (Z)–6–nonen–4–olide,
extractions were similar. However, some (Z)–5–octen–4–oilde, piperonal, tuberoholoside
chemicals in the tuberose absolutes from both and tuberolide (Nuntavan, 1996). Reverchon and
solvents were different; methyl anthranilate, 1- Poletto (1996) reported the main chemical
tetradecene and (Z)-methyl isoeugenol were components of tuberose absolute from super
detected only in the petroleum ether absolute, critical fluid extraction were: 1,8-cineole, methyl
while (Z)-3-hexenyl 2-oxopropanoate, 2,4,-di-tert- benzoate, methyl salicilate, trans-methyl eugenol
butylphenol and ecosanol were detected only in and benzyl benzoate. Moreover, Jumras and
the hexane absolute. Several studies have shown Possom (2003) reported tuberose oil chemical as
that the tuberose absolute contained many follows: methyl benzoate, methyl anthranilate,
chemical constituents, such as: benzyl benzoate, benzyl alcohol, butyric acid, eugenol, nerol,
(Z)–5–decen–4-oilde, (Z,Z)–6,9–dodecadien–4– farnesol and geraniol.
210 Kasetsart J. (Nat. Sci.) 43(5)

Figure 6 Chromatogram of tuberose absolute obtained from petroleum ether extraction.

Table 6 Chemical compositions of tuberose absolute obtained from petroleum ether extraction.
Peak No. Retention time (min) %Relative peak area Possible compounds
1 15.297 4.15 methyl anthranilate
2 15.741 0.98 1-tetradecene
3 16.022 1.89 methyl eugenol
4 16.757 4.85 methyl isoeugenol
5 17.117 0.96 (Z)-methyl isoeugenol
6 17.311 6.05 (E)-methyl isoeugenol
7 17.385 18.13 7-decen-5-olide
8 18.463 1.69 1-hexadecene
9 20.754 4.06 alpha-farnesol
10 21.935 10.28 benzyl benzoate
11 24.217 1.34 benzyl salicylate
12 32.462 3.65 tricosane
13 37.013 29.44 pentacosane
14 43.597 12.53 heptacosane

CONCLUSIONS ACKNOWLEDGEMENT

The percentage yield of absolutes This work was financially supported by

obtained from cold enfleurage, hot enfleurage, Kasetsart University Research and Development
hexane and petroleum ether extractions were Institute (KURDI) and Kasetsart Agricultural and
0.3137%, 6.5808%, 0.0279%, and 0.0182%, Agro-Industrial Product Improvement Institute
respectively. The main chemical component (KAPI).
detected in absolutes extracted by enfleurage was
methyl benzoate, while benzyl benzoate and LITERATURE CITED
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benzyl benzoate, 7-decen-5-olide and methyl plant volatile emission. Curr. Opin. Plant
salicylate, respectively. Biol. 8: 113-118.
 Kasetsart J. (Nat. Sci.) 43(5) 211

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