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Article

Supercritical CO2 Extraction of Narcissus poeticus L. Flowers for the Isolation of Volatile Fragrance Compounds

by
Renata Baranauskienė
and
Petras Rimantas Venskutonis
*
Department of Food Science and Technology, Kaunas University of Technology, LT-50254 Kaunas, Lithuania
*
Author to whom correspondence should be addressed.
Molecules 2022, 27(2), 353; https://doi.org/10.3390/molecules27020353
Submission received: 10 December 2021 / Revised: 30 December 2021 / Accepted: 1 January 2022 / Published: 6 January 2022
(This article belongs to the Special Issue Featured Papers on Bioactive Flavour and Fragrance Compounds 2022)

Abstract

:
The flowers of Narcissus poeticus are used for the isolation of valuable fragrance substances. So far, as the majority of these substances consist of volatile and sensitive to heat compounds, there is a need of developing effective methods for their recovery. In this study, freeze-dried N. poeticus inflorescences were extracted with pure supercritical CO2 (SFE-CO2) and its mixture with 5% co-solvent ethanol (EtOH) at 40 °C. Extract yields varied from 1.63% (12 MPa) to 3.12% (48 MPa, 5% EtOH). In total, 116 volatile compounds were identified by GC-TOF/MS in the extracts, which were divided into 20 different groups. Benzyl benzoate (9.44–10.22%), benzyl linoleate (1.72–2.17%) and benzyl alcohol (0.18–1.00%) were the major volatiles among aromatic compounds. The amount of the recovered benzyl benzoate in N. poeticus SFE-CO2 extracts varied from 58.98 ± 2.61 (24 MPa) to 91.52 ± 1.36 (48 MPa) mg/kg plant dry weight (pdw). α-Terpineol dominated among oxygenated monoterpenes (1.08–3.42%); its yield was from 9.25 ± 0.63 (12 MPa) to 29.88 ± 1.25 (48 MPa/EtOH) mg/kg pdw. Limonene was the major monoterpene hydrocarbon; (3E)-hexenol and heneicosanol dominated among alcohols and phenols; dihydroactinidiolide and 4,8,12,16-tetramethyl heptadecan-4-olide were the most abundant lactones; heptanal, nonanal, (2E,4E)-decadienal and octadecanal were the most abundant aldehydes. The most important prenol lipids were triterpenoid squalene, from 0.86 ± 0.10 (24 MPa) to 7.73 ± 0.18 (48 MPa/EtOH) mg/kg pdw and D-α-tocopherol, from 1.20 ± 0.04 (12 MPa) to 15.39 ± 0.31 (48 MPa/EtOH) mg/kg pdw. Aliphatic hydrocarbons (waxes) constituted the main part (41.47 to 54.93%) in the extracts; while in case of a 5% EtOH the percentage of alkanes was the lowest. The fraction of waxes may be removed for the separation of higher value fragrance materials. In general, the results obtained are promising for a wider application of SFE-CO2 for the recovery of fragrance substances from N. poeticus flowers.

1. Introduction

Aromatic herbs and flowers are an important source of natural ingredients for the development of flavours and fragrances. Narcissus is a genus of perennial spring flowering plants of the Monocotyledon family Amaryllidaceae. Carl Linnaeus originally defined six species of Narcissus in 1753, while N. poeticus was the first one described in his book Species Plantarum [1]. Nowadays the genus is generally considered to consist of about 12 sections with approximately 80 species. Poetic, commonly also called daffodil (N. poeticus), supposed (N. pseudonarcissus), tripod (N. triandrus), and bouquet (N. tazetta) narcissus are the most widely grown species, while cyclamen (N. cyclamineus), chaste (N. papyraceus), and fragrant (Narcissus × odorus) are less popular. Three species, namely white (N. poeticus), bouquet (N. tazetta) and yellow/supposed (N. pseudonarcissus) are most popular in Lithuania as ornamental plants in flower gardens, which also grow in old parks and meadows.
N. poeticus is a wild, bulbous herbaceous plant, growing up to 20–40 cm height. The leaves are radical, green, narrow and long; it blooms in late spring (April–May) with an extremely fragrant white flower per stem [2]. N. poeticus is thought to originate from the Middle East or Eastern Mediterranean; now it is spread all over Europe, particularly France, Spain, southern Italy and north-western Greece [3]. It is also naturalized in New Zealand, British Columbia, Canada, United States, North Africa, and Asia.
The plants of the Amaryllidaceae family have also been used as herbal remedies since ancient times. The Narcissus genus is a rich source of potentially valuable pharmaceuticals, such as alkaloids galanthamine, lycorine, isoquinolines narciclasine, pancratistatin, (E)-dihydronarciclasine, and their corresponding 7-deoxyanalogues, narcipavline, norlycoramine; some of them might be promising in the treatment of Alzheimer’s and oncological diseases [4,5,6,7,8]. Health beneficial chlorogenic (755.93 ± 4.06 μg/g), p-coumaric, ferulic acids and flavonoids (hyperoside, isoquercitrin, quercitrin) were reported in N. poeticus ethanolic extract from Romania [9].
Essential oils and extracts used in the perfumery industry are mostly produced from N. jonquilla, N. poeticus, N. tazetta and some other species. N. poeticus flowers as exceptionally fragrant material are cultivated in the Netherlands and southern France mainly for distilling essential oil (EO). Groom [10] described the aroma of the narcissus oil as resembling the combination of jasmine and hyacinth; therefore, its floral concrete or absolute has been used as a principal ingredient in ~11% of modern quality perfumes, including such famous names as “Fatale” or “Samsara”. The absolute of narcissus flowers has been widely used not only in popular classical perfumes in combination with rose, jasmine, violet and etc., but also in sophisticated modern perfumes, especially for strong pungent, green, woody and deep floral scent, which blends well with many other floral absolutes, such as ylang-ylang, jasmine, neroli, mimosa, clove bud carnation and others [2,11].
Industrially the absolutes are produced by extraction with organic solvent (hexane or other), its evaporation (causing some loss of volatile compounds), and removal of waxes. The distillation of fresh daffodil flowers recovers pure fraction of volatiles (essential oil), which are free of larger molecular weight waxes. However, the yields of EO are very low; for example, only 0.05% [12], while there is a risk of thermal degradation of some volatile compounds [11]. Nowadays green technologies for the isolation of valuable substances from plant materials are highly preferable in terms of safety and environmental protection; thus, instead of traditional distillation and/or solid-liquid extraction methods, the supercritical fluid extraction with carbon dioxide (SFE-CO2) was applied in our study for the recovery of lipophilic fraction containing fragrant volatile constituents. SFE-CO2 has been widely used for the recovery of EO together with other lipophilic constituents [13]. SFE-CO2 possesses many advantages (nontoxic, non-flammable, inexpensive, extraction at low temperatures avoids thermal degradation of the compounds and yields high purity extracts) and therefore can be successfully explored in pharmaceutical, cosmetics, food and other industries for recovery of valuable lipophilic constituents. On the other hand, the disadvantage of SFE-CO2 in the isolation of EO is that the extract also contains non-volatile lipophilic constituents.
More than 430 volatile compounds have been reported in Narcissus spp. [12,14,15,16,17,18,19]. For instance, 103 and 66 compounds were identified in N. trevithian and N. geranium oil, respectively, while only 20 of them were common for both species [17]. In the plants from Greece (E)-ocimene (61.12%) was dominant for N. tazetta and benzyl acetate (19.36%) for N. serotinus among 19 identified in the EOs constituents [12]. The main volatiles of N. tazetta absolute from Italy were γ-terpinene (27.21%), methyl cinnamate (15.84%), benzyl acetate (9.58%), benzyl alcohol (4.79%) and benzyl benzoate (4.00%) [18]. Eleven compounds were identified in the headspace of narcissus flowers; benzyl acetate (44.0%), 3,4- and 3,5-dimethoxy toluene (10.0 and 25.0%), and indole (5.0%) were major in the living flowers, whereas the percentages of the same compounds in the picked flowers were 30–43%, 18–39.5% and 0.3–1.0% [15]. (E)-β-Ocimene was found in high percentages in six species of Narcissus grown in Spain [19]. The main constituents of the Chinese narcissus flowers were (E)-β-ocimene (62.73–66.06%), benzyl acetate (11.65–25.02%), (Z)-β-ocimene, 1,8-cineole, and linalool [20].
Volatile compounds of N. poeticus have also been studied [2,11,21,22]. Ehret et al. [22] identified 80 new minor volatile components in N. poeticus absolute from France “Massif central”, α-terpineol (23.7%), methyl (E)-isoeugenol (20.0%), and benzyl benzoate (19.4%) being the major ones; more than 20 of them were considered as responsible for the complex floral notes reminiscent of “rose”, “jasmine”, “violet”, “tuberose” and “orange flower”. Cinnamyl alcohol (29.91 μg/g), methyl isoeugenol (28.07 μg/g), isoeugenol (23.12 μg/g), methyl eugenol (20.72 μg/g), and α-terpineol (20.31 μg/g) were the main components in hexane extract of fresh N. poeticus flowers of Rocca di Mezzo [11], whereas cinnamyl alcohol (30.2 μg/g), benzyl benzoate (28.5 μg/g), isoeugenol methyl ether (28.1 μg/g), (Z)-ocimene (25.2 μg/g), isoeugenol (22.9 μg/g) were quantitatively major in the plants from Sirente-Velino (Abruzzo region, Italy) [2]. To the best of our knowledge, only one article is available on SFE-CO2 of N. poeticus [11]; however, this study applied comparatively low pressure of the fluid CO2 (12 MPa). So far, as the solubility of many compounds increases with increasing solvent density, which depends on its pressure, in our study we applied CO2 pressure up to 48 MPa. It is important to mention that some of these volatiles are stated as more or less allergenic compounds, for example, cinnamyl alcohol, isoeugenol and cinnamyl aldehyde, and their use must follow legislation. It was suggested that in the preparation of safer perfumes absolute can be obtained from the corona only instead of tepals plus corona [2].
Considering previously reported data, our study aimed at evaluating the effects of pressure and addition of co-solvent in SFE-CO2 of freeze-dried N. poeticus flowers. Extract yields and comprehensive analysis of the recovered volatiles compounds were performed for this purpose. It is expected that the data obtained will expand our knowledge on the composition of N. poeticus fragrant constituents and the possibilities of their recovery by using green extraction techniques.

2. Results and Discussion

2.1. Extract Yields

The yields of the SFE-CO2 extracts of N. poeticus freeze-dried flowers varied from 1.63 ± 0.29 to 3.12 ± 0.12% (Table 1). The temperature of 40 °C, static extraction time 10 min and dynamic extraction time of 120 min were kept constant in all experiments. According to the literature reports, the yields of the concrete (absolute + waxes) obtained with hexane extraction were 0.2–0.3% [22], 0.41–0.45% [2,11] of fresh flowers and with petroleum ether –0.2% of fresh flowers [23]. The percentages of absolute in the concretes varied from 27 to 37% [11,23].
There were no statistical differences in the extract yields at 12–36 MPa, while at 48 MPa the yield of the recovered fraction was significantly higher (p < 0.05). Additionally, 5% ethanol applied as a co-solvent increased extract yield by ~32% (from 2.36 to 3.12%) compared with pure SFE-CO2 at 48 MPa. Ferri et al. [11] compared conventional hexane extraction with SFE-CO2 of N. poeticus; however, the absolute yields of SFE-CO2 extracts are not available in their article. Nevertheless, it is evident that SFE-CO2 extracts contained all the main constituents, which were identified in hexane extracts, while their yields in SFE-CO2 were significantly lower, even in case of using 5% ethanol, which enhanced the yields of most characteristics compounds [11].

2.2. Composition of N. poeticus SFE-CO2 Extracts

The detailed list of extracted by SFE-CO2 volatile compounds, their percentage composition and odour descriptions are presented in Table 2. In total, 116 compounds were identified in the SFE-CO2 extracts of N. poeticus flowers obtained under different extraction conditions. The sums (in %) and the numbers of the identified compounds in SFE-CO2 extracts at 12 MPa, 24 MPa, 36 MPa, 48 MPa and 48 MPa/EtOH were 77.47/105, 74.12/100, 76.07/104, 74.10/93, and 76.69/92, respectively.
It is evident that the composition of N. poeticus SFE-CO2 extracts is very complex, therefore for further discussion all the identified compounds were divided into 20 different classes, namely aromatics, aliphatic hydrocarbons, aromatic hydrocarbons, monoterpene hydrocarbons, oxygenated monoterpenes, oxygenated sesquiterpenes, esters, alcohols, aldehydes, ketones, lactones, acids, amides, diterpenoids, triterpenoids, tocopherols, phenylpropanoids, phenols and others (including oxanes, heteroaromatics, etc) (Table 2).

2.2.1. Aromatics

Aromatic compounds is the most important group of volatiles in N. poeticus SFE-CO2 extracts; their percentage content varied from 12.55% (24 MPa) to 14.32% (48 MPa/EtOH). The major compounds were benzyl benzoate (65, 9.44–10.22%), benzyl linoleate (107, 1.72–2.17%), benzyl alcohol (12, 0.18–1.00%) and benzyl linolenate (108, 0.26–0.53%). Other aromatic compounds include hydrocinnamyl alcohol (30), 2-phenylethyl benzoate (70), benzyl 4-methoxybenzoate (75), benzoic acid hexadecyl ester (104), benzoic acid (21), benzyl oleate (106) (<0.1–0.2%), while hexyl benzoate (60) and (3Z)-hexenyl benzoate (58) were detected in trace amounts (Table 2).
Benzyl benzoate, benzyl alcohol and benzoic acid are used in a wide variety of cosmetics formulations as fragrance ingredients and preservatives. The group of benzyl derivatives was reaffirmed as generally recognized as safe (GRAS) by the Expert Panel of the Flavour and Extract Manufacturers (FEMA), and the evidence of safety is supported by the fact that the intake of benzyl derivatives as natural components of traditional foods is larger than their intake in the case of intentional adding as flavouring substances [24]. Benzyl alcohol is an aromatic alcohol, which has been characterised as possessing “sweet, flower” [25] and “berry, cherry, grapefruit, citrus, and walnut” [26] odour notes. The absolute amounts of main volatile compounds in mg/kg pdw are summarised in Figure 1 and Figure 2. The highest recoveries of benzyl alcohol (12, 8.79 ± 0.59 mg/kg pdw), as well as benzyl linoleate (107, 18.96 ± 0.42 mg kg pdw) were obtained at 48 MPa/EtOH; however this value was not significantly different to the one obtained at 48 MPa (Figure 1A). Benzyl benzoate and benzyl alcohol were previously reported in high amounts in the absolutes of hexane extracts of N. poeticus from Italy [2,11].
It was reported that narcissus absolute has “a very strong, green, earthy and woody” odour, and that in appropriate dilution, releases a characteristic blend of floral and balsamic notes: light floral notes (rose or jasmine), deep floral notes (ylang-ylang, tuberose, orris and violet), notes of the balsamic type (styrax) and woody, earthy scents (oakmoss or patchouli) [22]. The balsamic type (styrax) flavour notes can be associated with the presence of benzoates and cinnamates in the N. poeticus SFE-CO2 products composition. Benzyl benzoate also possesses “balsamic, oil, herb” [25] and “almond, cheese, cherry, floral, pineapple, strawberry, sweet” [26] aroma notes. The absolute amount of the recovered benzyl benzoate from N. poeticus by SFE-CO2 varied from 58.98 ± 2.61 mg/kg pdw (24 MPa) to 91.52 ± 1.36 mg/kg pdw (48 MPa) (Figure 1A).
It was also reported that benzyl benzoate, its derivatives and benzyl cinnamate may be promising compounds in reducing hypertension [28]. It may be concluded that N. poeticus SFE-CO2 extracts after the separation of waxes might be a potential natural source of benzyl benzoate and benzyl alcohol.

2.2.2. Monoterpene Hydrocarbons, Oxygenated Monoterpenes and Sesquiterpenes

α-Terpineol (25), which possess “oil, anise, mint” [25] and “lilac” [26] aroma notes, was major constituent among oxygenated monoterpenes (1.08–3.42%); it was also reported previously in N. poeticus absolute in comparatively high percentages [2,11,22]. The absolute amount of the recovered α-terpineol in N. poeticus SFE-CO2 extracts increased by increasing pressure and varied from 9.25 ± 0.63 mg/kg pdw (12 MPa) to 29.88 ± 1.25 mg/kg pdw (48 MPa/EtOH) (Figure 1B). It is well known that the quantitatively minor and even trace components can be important on the overall odour quality of flavours and fragrances. Consequently, the presence of ketones β-ionone epoxyde (0.28–0.42%) and trace component β-ionone may be important even at low concentrations (≤0.04%) by providing “violet flower aroma notes” of narcissus.
Limonene was the major monoterpene hydrocarbon; however, its percentage was low (0.08–0.17%). It is interesting that (E)-β-ocimene was not found in SFE-CO2 extracts of N. poeticus from Lithuania, while this compound was reported to be present in six species of Narcissus from Spain [19], N. tazetta EOs from Greece and N. poeticus absolute from Italy [2].

2.2.3. Alcohols, Phenols, Lactones, Aldehydes and Other Volatiles

Among alcohols and phenols (3E)-hexenol (1) and heneicosanol (101) were recovered at the highest amounts (Figure 1C). (3E)-Hexenol is very abundant in various aromatic and spicy plants and possesses “moss” and “fresh” aroma notes. Therefore it may provide “green, woody” scents for narcissus even at low percentage concentrations, which varied from 0.10 ± 0.02% (36 MPa) to 0.89 ± 0.06% (48 MPa) (Table 2). The recovery of long-chain alcohol heneicosanol from N. poeticus by SFE-CO2 varied from 6.07 ± 0.30 mg/kg pdw (24 MPa) to 10.47 ± 0.62 mg/kg pdw (12 MPa) (Figure 1C).
The recoveries of esters (Figure 1D) and acids (Figure 2A) were quite low; however, it is interesting to mention that the amounts extracted from N. poeticus ethyl hexadecanoate (78), ethyl linolenate (90) and linoleic acid (86) significantly increased at 48 MPa and adding 5% of EtOH. It is believed that high amount of alcohols and long-chain acids in narcissus has more effect on the longer lasting trait of its odour, rather than on the odour quality [17].
Dihydroactinidiolide (57, 0.42–0.64%) and 4,8,12,16-tetramethyl heptadecan-4-olide (98, 0.27–0.50%) were the most abundant lactones in the SFE-CO2 extracts of N. poeticus (Figure 2B). The absolute content of dihydroactinidiolide was from 3.57 ± 0.23 mg/kg pdw (12 MPa) a to 5.70 ± 0.10 mg/kg pdw (48 MPa). Dihydroactinidiolide is a lactone (cyclic ester) resulting from the secondary oxidation of β-ionone through the intermediate 5,6-epoxy-β-ionone [29]. It possesses a “sweet, tea-like” odour and is used as a fragrance. Dihydroactinidiolide occurs naturally in plant leaves, black tea, fenugreek, fire ants, fruits, tobacco and it is a pheromone for a variety of insects [30]. Dihydroactinidiolide was reported to accumulate in Arabidopsis leaves under high light stress [31] and to exhibit cytotoxic effects against cancer cell lines [32].
Heptanal (2), nonanal (19), (2E,4E)-decadienal (37) and octadecanal (80) were the most abundant aldehydes (Figure 2C). Nonanal possesses very complex odour (Table 2) including “green” and “rose” scents. Its content in the extracts was 0.31–0.48%, while the absolute amounts, which were recovered from the flowers, varied from 2.70 ± 0.06 mg/kg pdw (48 MPa/EtOH) to 3.82 ± 0.16 mg/kg pdw (48 MPa) (Figure 2C). The amount of octadecanal was from 2.58 ± 0.18 mg/kg pdw (24 MPa) to 4.10 ± 0.22 mg/kg pdw (12 MPa). Octadecanal (or stearyl aldehyde) is a long chain fatty aldehyde and was identified as a biologically active pheromone component. Octadecanal is often used as the substrate of choice to test the microsomal enzyme fatty aldehyde dehydrogenase activity in patients suspected of having Sjogren-Larsson syndrome (autosomal recessively inherited neurocutaneous disorder) [33].

2.2.4. Triterpenoids, Tocopherols and Others

Supercritical CO2 together with volatile compounds also extracts higher molecular weight lipophilic compounds, among them important prenol lipids such as triterpenoid squalene and tocopherols, which are well known antioxidants. Squalene is a long chain triterpene hydrocarbon, which is a precursor in the synthesis of sterols. The amount of squalene varied from 0.86 ± 0.10 mg/kg pdw (24 MPa) to 7.73 ± 0.18 mg/kg pdw (48 MPa/EtOH). It is worth noting that the use of a co-solvent ethanol increased the amount of extracted squalene ~2.5 times (Figure 2D). The antioxidant and oxygen carrying properties of squalene predicts its potential use in preventing cardiovascular diseases and cancer [34].
The amount of the recovered vitamin E (D-α-tocopherol) was from 1.20 ± 0.04 mg/kg pdw (12 MPa) to 15.39 ± 0.31 mg/kg pdw (48 MPa/EtOH). In nature, there are four main structures of tocopherols, namely α-, β-, γ-, and δ-tocopherol. Tocopherols are lipophilic molecules which are synthesized by plant cells and stored in leaves and seeds, and are endowed with antioxidant functions. They possess multiple beneficial healthy effects, such as the prevention of cardiovascular diseases and cancer [35].
The amide of oleic acid (Z)-9-Octadecenamide (99) and amide of palmitic acid–hexadecanamide (91), which are fatty acid derivatives, were determined in the lipophilic fraction. The percentage of (Z)-9-octadecenamide (99) was from 0.44 to 1.00% (Table 2) and the absolute amount (Z)-9-octadecenamide varied from 3.25 ± 0.13 mg/kg pdw (24 MPa) to 8.76 ± 0.96 mg/kg pdw (48 MPa/EtOH) (Figure 2D). Oleamide was reported in various natural plant materials such as Nigella sativa [36], mountain celery seeds [37] and other plants [38], however it should be noted that it may be an artefact, which is transferred into the extracts from the labware [39]. The health-promoting properties of (Z)-9-octadecenamide have been reported for this oleamide, such as anti-inflammatory and antibacterial activities [36], hypolipidemic activity [37], against Alzheimer disease [38], and etc.
Some of the components depending on different classes were identified in narcissus extract when the highest pressure and addition of 5% ethanol as co-solvent were applied, e.g., pantolactone (13), ethyl heptanoate (17), benzoic acid (21), (E)-8-hydroxylinalool (40), 3-hydroxydecanoic acid (56), ethyl tetradecanoate (66), ethyl 9-octadecenoate (88).

2.2.5. Aliphatic Hydrocarbons (Waxes)

It was observed that the saturated hydrocarbons (n-alkanes) constitute a very large fraction in the total amount (from 41.47 to 54.93%) of GC-detectable volatiles of the narcissus lipophilic fraction (SFE-CO2 extracts). The percentage of n-alkanes slightly decreased by increasing pressure; it was expected additionally that by applying 5% ethanol as a co-solvent the percentage amount of alkanes was the lowest (41.47%). Higher n-alkanes, such as heptacosane (105, 11.21–15.40%), tricosane (96, 7.57–11.36%), nonacosane (111, 5.64–7.27%), untriacontane (115, 4.41–6.73%), pentacosane (102, 4.18–6.13%) and heneicosane (81, 3.71–5.50%) were the major qualitative compounds in this fraction (Table 2).
The highest absolute amounts of aliphatic hydrocarbons were extracted at 12 MPa, while the lowest at 24 MPa. Further increase in pressure from 24 to 48 MPa resulted in the increase in the recovered amounts of n-alkanes, while in the case of using ethanol it was reduced again. For example, the highest amount of the recovered n-heptacosane (105) was 131.49 ± 7.00 mg/kg pdw at 12 MPa and the lowest 84.07 ± 1.37 mg/kg pdw at 24 MPa (Figure 3). At further increase in pressure its amount increased to 121.88 ± 5.30 mg/kg pdw (48 MPa), while 5% ethanol reduced the value to 98.08 ± 5.30 mg/kg pdw, which was significantly lower to that extracted at 12 MPa. The amount of these saturated acyclic alkanes could be reduced by using conventional dewaxing procedures.

3. Materials and Methods

3.1. Plant Material and Chemicals

Narcissus poeticus L. plants were grown in a farmstead near the city of Klaipėda (Lithuania, coordinates: 55°45′ N 21°10′ E) and the flowers were collected at a full flowering stage. The flowers were picked in the morning manually, separated from the stems by scissors. Fresh flowers were frozen to −40 °C and freeze-dried in a Sublimator 40 at 0.05 mbar pressure (Zirbus Technology, Bad Grund, Germany). The dried flowers were ground in an ultra-centrifugal mill ZM 200 (Retsch, Haan, Germany) using 0.5 mm hole size sieve.
Pentane (for residue analysis, ≥99.0%) was from Sigma-Aldrich (Steinheim, Germany), Ethanol (≥96%) obtained from Joint Stock Company Stumbras, Lithuania. C7–C30 saturated n-alkanes standard and internal standard (decane) were from Supelco Analytical (Bellefonte, PA, USA). The following reference compounds (95–99% purity) for the identification of narcissus volatiles were purchased from Fluka, Sigma–Aldrich or Supelco: limonene, p-cymene, α-terpineol, vanillin, benzyl alcohol, nonanal, β-ionone, caryophyllene oxide, squalene.

3.2. Supercritical Carbon Dioxide Extraction (SFE-CO2)

SFE-CO2 was carried out in a supercritical fluid extractor Helix (Applied Separation, Allentown, PA, USA). Each extraction was performed from 10 g of ground N. poeticus flowers placed in a 50 mL cylindrical extractor vessel (14 mm × 320 mm; h/d = 22.86), between two layers of defatted cotton wool in both ends, to avoid particles clogging the system. The temperature of the extraction vessel was controlled by a surrounding heating jacket. The flow rate of CO2 in the system (v) was controlled manually by the micro-metering valve (back-pressure regulator). The volume of CO2 consumed was measured by a ball float rotameter and a digital mass flow meter in liters per min (SL/min) at standard state: pressure P = 100 kPa, temperature T = 20 °C, density ρ = 0.0018 g/mL. The conditions for extraction were set as follows (see in Table 1): extraction time 120 min, pressure 12–48 MPa, extraction temperature 40 °C, flow rate of CO2 2 L/min. A static time of 10 min was included in to the total extraction time and was constant. The extraction at 48 Mpa was also performed using 5% (v/v) ethanol as a co-solvent in order to enhance the polarity of the solvent mixture. The extracts were collected in brown glass bottles at room temperature and atmospheric pressure and stored at −20 °C until further analysis.

3.3. Gas Chromatography–Time-of-Flight Mass Spectrometry (GC-TOF/MS)

For qualitative and quantitative composition of SFE-CO2 extracts of N. poeticus flowers, they diluted in pentane (10 mg/mL) containing 0.2% internal standard and analysed on a GC × GC-TOFMS LECO Pegasus 4D system, consisting of an Agilent 7890A GC, a GERSTEL Multipurpose Sampler MPS (Gerstel GmbH, Mulheim an der Ruhr, Germany), a high-speed TOF/MS detector (LECO, St. Joseph, MI, USA) and a four jet cryogenic modulator (Zoex, Houston, TX, USA) by comparing the 1D first dimension linear temperature programmed Kováts retention index with the peak identities provided by a mass spectral similarity search. The column set consisted of a primary column BPX-5 (30 m, 0.25 mm i.d., 0.25 μm film thickness) (SGE Analytical Science, Australia) connected to a secondary column, BPX-50 (1.8 m, 0.10 mm i.d., 0.1 μm film thickness). The primary oven was programmed as follows: 50 °C ramped to 100 °C at 10 °C/min (0 min) further ramped to 300 °C at 5 °C/min (hold 5 min); the secondary oven programming was from 65 °C ramped to 115 °C at 10 °C/min (0 min) then ramped to 315 °C at 5°C/min (hold 5 min). The transfer line temperature was 250 °C, the GC injector port was kept at 280 °C with desorption time of 5 min.
The TOF/MS acquisition rate was 10 spectra/s, the mass range used for identification was from 35–550 m/z units. Detector’s voltage was set at 1550 V and ion source temperature of 250 °C. Data from the GC × GC-TOF/MS system were collected by ChromaTOF software v.4.22 (LECO) after a solvent peak delay of 420 s in a splitless mode for 30 s, a further valve was opened and purge flow was 20 mL/min; mass spectrum assignment was based on matching against Adams, NIST, MainLib, RepLib mass spectra libraries; signal-to-noise threshold was set as 50 with the minimum similarity accepted was 700. The mean values were calculated from triplicate injections.
Quantitative data were obtained by peak area normalization without using correction factors as the means of triplicate GC-TOF/MS runs and expressed as peak area percentage and recalculated in mg/kg per plant dry weight (mg/kg pdw). Quantitative data were calculated using the formula: m i = m D × A i / A D × R F i × Y e × 1000 / 100 / m e , where m i is the mass of the individual compound i to be quantified, expressed in mg/kg per plant dry weight (mg/kg pdw); m D mass of decane (internal standard, IS), A i and A D —are the peak area of the analyte i and the IS, respectively, R F i —response factor of individual compound; Y e —the yield of the extract (%), m e —the mass of the extract (g).
The identification of volatile components was assigned by comparing their Kováts Retention Indices (KI) relative to C7–C30 n-alkanes, obtained on nonpolar BPX-5 column with those provided in literature [27] and by comparing their mass spectra with the data provided by the NIST, MainLib, RepLib and Adams mass spectral libraries. The identity of some constituents was confirmed by co-injection of reference compounds. Positive identification was assumed when good match of mass spectrum and KI was achieved.

3.4. Statistical Analysis

All analyses were replicated at least three times and all data are reported as mean values ± standard deviations (SD) using MS Excel 2010 software. Data were statistically handled by one-way analysis of variance (ANOVA, vers. 2.2), significant differences among the samples were evaluated by the Duncan’s multiple-range test at the probability level of p < 0.05.

4. Conclusions

Narcissus poeticus flowers were extracted with supercritical carbon dioxide at different solvent pressures, from 12 to 48 MPa. The yield of lipophilic fraction significantly increased after raising the pressure from 36 to 48 MPa and by adding 5% of a co-solvent ethanol into the CO2 flow. Generally, the yields were higher compared with the previously reported data for conventional extraction with organic solvents. In total, 116 volatile compounds were identified by GC-TOF/MS in the extracts. The most important for N. poeticus odour constituents benzyl benzoate (9.44–10.22%), benzyl linoleate (1.72–2.17%) and benzyl alcohol (0.18–1.00%) were the major volatiles among aromatic compounds. The extracts contained a large fraction of waxes, which are not desirable in the production of higher quality fragrance ingredients; however, adding a co-solvents ethanol enabled the reduction in the percentage of higher alkanes, while the amount of the recovered benzyl aromatics increased. On the other hand, for practical purposes, SFE-CO2 at 48 MPa would most likely be preferable because in this case the process becomes less complicated and cheaper; there is no need for a co-solvent pump and removal of it after extraction, which may result in some loss of volatile fragrance constituents. However, adding co-solvent increases the recovery of an important bioactive compound α-tocopherol approx. 2-fold. In general, the results obtained are promising for a wider application of supercritical extraction for the recovery of fragrance substances from Narcissus poeticus flowers.

Author Contributions

Conceptualization, P.R.V.; methodology, R.B.; software, P.R.V. and R.B.; investigation, R.B.; resources, P.R.V.; writing—original draft preparation, R.B. and editing P.R.V.; visualization, R.B.; supervision, P.R.V.; project administration, P.R.V. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data available from the authors upon request.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. The absolute amounts of the main volatile compounds of major classes ((A)—Aromatics, (B)—Oxygenated monoterpenes and sesquiterpenes, (C)—Alcohols and phenols, (D)—Esters) of Narcissus poeticus SFE-CO2 extracts, in mg/kg pdw. Constituents are numbered by the same order as provided in Table 2. Values within columns followed by the same letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
Figure 1. The absolute amounts of the main volatile compounds of major classes ((A)—Aromatics, (B)—Oxygenated monoterpenes and sesquiterpenes, (C)—Alcohols and phenols, (D)—Esters) of Narcissus poeticus SFE-CO2 extracts, in mg/kg pdw. Constituents are numbered by the same order as provided in Table 2. Values within columns followed by the same letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
Molecules 27 00353 g001
Figure 2. The absolute amounts of main volatile compounds of major classes ((A)—Acids, (B)—Lactones, (C)—Aldehydes, (D)—Amides and prenol lipids) of Narcissus poeticus SFE-CO2 extracts, in mg/kg pdw. Constituents are numbered by the same order as provided in Table 2. Values within columns followed by the same letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
Figure 2. The absolute amounts of main volatile compounds of major classes ((A)—Acids, (B)—Lactones, (C)—Aldehydes, (D)—Amides and prenol lipids) of Narcissus poeticus SFE-CO2 extracts, in mg/kg pdw. Constituents are numbered by the same order as provided in Table 2. Values within columns followed by the same letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
Molecules 27 00353 g002
Figure 3. The absolute amounts of aliphatic hydrocarbons in Narcissus poeticus SFE-CO2 extracts, in mg/kg pdw. Constituents are numbered by the same order as provided in Table 2. Values within columns followed by the same letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
Figure 3. The absolute amounts of aliphatic hydrocarbons in Narcissus poeticus SFE-CO2 extracts, in mg/kg pdw. Constituents are numbered by the same order as provided in Table 2. Values within columns followed by the same letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
Molecules 27 00353 g003
Table 1. SFE-CO2 extraction conditions of Narcissus poeticus and obtained yields (%).
Table 1. SFE-CO2 extraction conditions of Narcissus poeticus and obtained yields (%).
SFE-CO2 Extraction ConditionsExtract Yield, %
12 MPa1.63 ± 0.29 a
24 MPa1.75 ± 0.10 a
36 MPa1.82 ± 0.09 a
48 MPa2.36 ± 0.09 b
48 MPa + 5% EtOH3.12 ± 0.12 c
Results are expressed as a mean ± standard deviation (n = 3); Values within columns followed by the same letter (a–c) do not differ statistically at p < 0.05 (Duncan test).
Table 2. Chemical composition of Narcissus poeticus SFE-CO2 extracts, peak area percentage (%).
Table 2. Chemical composition of Narcissus poeticus SFE-CO2 extracts, peak area percentage (%).
No.#Compound AKI
Calc. B
KI
Lit. C
KI
Lit. D
12 Mpa24 Mpa36 Mpa48 Mpa48 Mpa/EtOHOdour Description 1
1.(3E)-Hexenol85485313860.35 ± 0.02 c0.67 ± 0.10 b0.10 ± 0.02 a0.89 ± 0.06 d0.72 ± 0.06 bmoss, fresh
2.Heptanal90790211740.20 ± 0.01 c0.27 ± 0.02 d0.26 ± 0.01 b0.24 ± 0.02 b0.15 ± 0.01 afat, citrus, rancid 1; oily, fruity, woody, fatty, nutty 2
3.tetrahydro-Citronellene938937 tr a0.05 ± 0.00 btr atr a0.07 ± 0.00 c
4.2-methyl-Nonane971970 E961 F0.06 ± 0.00 b0.09 ± 0.01 e0.07 ± 0.01 c0.08 ± 0.00 dtr a
5.Hexanoic acid97897318290.24 ± 0.02 a0.47 ± 0.05 d0.45 ± 0.03 b0.39 ± 0.00 e0.45 ± 0.04 bcdsweat 1; cheese, fatty, sour 2
6.2-Pentyl furan99298812400.13 ± 0.00 a0.22 ± 0.01 d0.19 ± 0.01 c0.16 ± 0.01 b green bean, butter 1; green, vegetable 2
7.Mesitylene9989951220 Etr atr atr a0.18 ± 0.00 b
8.Decane *1000100010000.12 ± 0.01 a0.15 ± 0.00 b0.17 ± 0.01 b0.20 ± 0.02 d0.20 ± 0.01 dalkane
9.(2E,4E)-Heptadienal1014100714010.10 ± 0.00 a0.30 ± 0.04 c0.15 ± 0.03 b nut, fat 1; cinnamon, hazelnut, fatty 2
10.p-Cymene102510251261tr atr btr atr a solvent, gasoline, citrus
11.Limonene1028102911780.12 ± 0.00 b0.17 ± 0.01 d0.15 ± 0.00 c0.11 ± 0.01 b0.08 ± 0.00 alemon, orange 1; citrus, sweet 2
12.Benzyl alcohol1034103118650.18 ± 0.01 a0.77 ± 0.04 b0.84 ± 0.05 b0.96 ± 0.01 c1.00 ± 0.02 csweet, flower 1; berry, cherry, grapefruit, citrus, walnut 2
13.Pantolactone10371032 F2034 F 0.06 ± 0.00cotton candy
14.Heptanoic acid10781074 E1918 E0.40 ± 0.06 a0.51 ± 0.05 c0.52 ± 0.05 bc0.47 ± 0.06 b0.60 ± 0.02 dsour 2
15.p-Cresol108310762067tr a0.06 ± 0.00 btr c0.06 ± 0.00 b0.10 ± 0.01 dmedicine, phenol, smoke 1; woody, ethereal, medicinal 2
16.2-Nonanone109710901388tr atr atr a hot milk, soap, green 1; cheese, coconut, oily, fatty, herbaceous, floral, fruity, fishy, soapy, waxy 2
17.Ethyl heptanoate11021093 E1321 E tr
18.(4E)-Nonenal11061096 G1458 Etr atr btr ab berry, melon, peach, pineapple, plum 2
19.Nonanal1109110013850.33 ± 0.01 c0.48 ± 0.02 d0.44 ± 0.01 b0.43 ± 0.04 b0.31 ± 0.01 afat, citrus, green 1; apple, coconut, grape, lemon, grapefruit, lime, melon, orange, nutty, citrus, oily, waxy, fatty, peach, rose, vegetable, fishy, meaty 2
20.(2E)-Nonenal116711611527 tr cucumber, fat, green 1; waxy, fatty 2
21.Benzoic acid11631162 E1624 0.11 ± 0.02urine 1; balsam 2
22.δ-Terpineol117011661655tr atr abtr ab0.05 ± 0.00 b0.06 ± 0.00 c
23.Octanoic acid1183117120830.05 ± 0.00 a0.07 ± 0.02 c0.06 ± 0.0 abc0.05 ± 0.00 a sweat, cheese 1; oily 2
24.Naphthalene119111811712 Etr abtr abtr b
25.α-Terpineol1194118816881.08 ± 0.04 a2.07 ± 0.10 b2.12 ± 0.06 b2.52 ± 0.19 c3.42 ± 0.16 doil, anise, mint 1; lilac 2
26.(2E)-Hexenyl butanoate119711941458 Etr atr a0.08 ± 0.00 b apple, cheese, green, meaty 2
27.n-Dodecane120012001200 tr abtr abtr b alkane
28.exo-2-Hydroxycineole12191219 E1845 E tr abtr abtr atr b
29.2-Hydroxycineole12301229 E 0.06 ± 0.01 a0.11 ± 0.02 b0.08 ± 0.00 a0.08 ± 0.01 a0.11 ± 0.00 b
30.Hydrocinnamyl alcohol12421231 E1989 Etr b0.09 ± 0.00 c0.05 ± 0.00 btr a0.09 ± 0.01 cbalsam, hyacinth, floral, sweet 2
31.Carvone1249124317200.05 ± 0.00 b0.08 ± 0.01 d0.06 ± 0.01 ctr atr acaraway, mint, basil, fennel 1; herbaceous 2
32.Nonanoic acid1280127022020.23 ± 0.01 a0.26 ± 0.03 bc0.26 ± 0.01 bc0.23 ± 0.03 a0.27 ± 0.02 cgreen, fat 1; cheese, waxy 2
33.n-Tridecane130113001300tr a0.06 ± 0.00 bc0.06 ± 0.00 b0.06 ± 0.01 bc0.06 ± 0.00 calkane
348-Hydroxymenthol130413012167 E0.06 ± 0.01 a0.05 ± 0.01 a0.05 ± 0.00 a0.06 ± 0.01 a0.10 ± 0.01 b
35.(2E,4Z)-Decadienal13061303 E17100.17 ± 0.01 b0.44 ± 0.02 e0.28 ± 0.0 c0.21 ± 0.02 dtr afried, fat
36.Undecanal130813062444 0.07 ± 0.01 btr a0.05 ± 0.01 ab oil, pungent, sweet
37.(2E,4E)-Decadienal1317131617100.27 ± 0.01 b0.68 ± 0.04 e0.45 ± 0.00 d0.32 ± 0.03 ctr afried, wax, fat 1; fatty, citrus, meaty 2
38.(2E)-Undecenol136613651899 F0.07 ± 0.00 c0.09 ± 0.00 b0.09 ± 0.00 b0.05 ± 0.01 a
39.Decanoic acid1367136623610.17 ± 0.00 b0.26 ± 0.03 d0.24 ± 0.02 c0.13 ± 0.01 a rancid, fat 1; fatty, citrus 2
40.(E)-8-Hydroxylinalool137013672265 E 0.13 ± 0.00
41.(E)-p-Menth-6-en-2,8-diol137713741740 Etr atr abtr a0.05 ± 0.00 b0.12 ± 0.01 c
42.3-Dodecanone139113901655 E0.05 ± 0.01 a0.05 ± 0.00 a0.06 ± 0.00 a0.06 ± 0.01 a0.11 ± 0.00 b
43.n-Tetradecane140014001400tr ctr atr b alkane
44.Vanillin140614072540 Etr a 0.08 ± 0.01 bcaramel, chocolate, sweet, vanilla 2
45.Carvone hydrate143214231754 Etr a tr atr abtr b
46.1b,5,5,6a-Tetramethyl-octahydro-1-oxa-cyclopropa[a]inden-6-one14541445 G tr atr b0.11 ± 0.01 ctr ab
47.(E)-Isoeugenol145514512372tr atr b 0.13 ± 0.01 cflower 1; clove, sweet, woody, spicy 2
48.3a,4,7,7a-tetrahydro-3a-methyl-2(3H)-Benzofuranone14571456 E2235 Etr atr btr abtr ab0.13 ± 0.01 c
49.(E)-β-Ionone148914881912tr abtr ctr bctr abtr aseaweed, violet, flower, raspberry 1; almond, basam, berry, grape, jam, orange, fruity, woody, floral, peach, raspberry, rose, sweet, violet, minty, wine-like, vegetable 2
50.β-Ionone epoxide14941488 E1957 E0.28 ± 0.01 a0.42 ± 0.02 d0.40 ± 0.01 b0.41 ± 0.02 c0.40 ± 0.02 bcfruit, sweet, wood
51.(E)-Methyl isoeugenol149714922176 Etr atr btr btr b0.05 ± 0.00 cspicy 2
52.2-Tridecanone149914961805 Etr a0.05 ± 0.00 b0.05 ± 0.00 b0.05 ± 0.00 btr aspicy 1; herbaceous 2
53.n-Pentadecane1500150015000.05 ± 0.01 btr a0.07 ± 0.00 c0.06 ± 0.00 bc0.06 ± 0.00 bcalkane
54.Tridecanal151515101824tr abtr atr ab 0.05 ± 0.01 cflower, sweet, must
55.Methyl dodecanoate153015251795tr atr btr btr b0.06 ± 0.00cfat, coconut 1; coconut, creamy, soapy, waxy 2
56.3-Hydroxydecanoic acid15361534 0.10 ± 0.01
57.Dihydroactinidiolide15461535 E2308 E0.42 ± 0.01 a0.63 ± 0.03 d0.62 ± 0.01 bc0.64 ± 0.04 cd0.62 ± 0.02 b
58.(3Z)-Hexenyl benzoate157015662122 E0.05 ± 0.00 b tr a tr awoody, herbaceous, green 2
59.γ-Undecalactone157615702270tr bctr btr btr btr aapricot 1; musty, peach, sweet, earthy 2
60.Hexyl benzoate158215802066 Etr balsam, woody, green 2
61.Caryophyllene oxide158715831962tr a tr b herb, sweet, spice
62.γ-Dodecalactone1687167723840.12 ± 0.00 b0.13 ± 0.01 bc0.14 ± 0.00 c0.14 ± 0.01 bc0.09 ± 0.00 afruit, sweet 1; musty, fatty, fruity 2
63.2-Pentadecanone169716972016 E0.17 ± 0.00 a0.17 ± 0.01 ab0.18 ± 0.00 ab0.18 ± 0.01 b0.16 ± 0.00 a
64.Heptadecane170017001700tr btr ctr btr a alkane
65.Benzyl benzoate1763176020719.90 ± 0.34 a9.44 ± 0.44 a10.22 ± 0.16 ab10.22 ± 0.69 b9.85 ± 0.12 abalsamic, oil, herb 1; almond, cheese, cherry, floral, pineapple, strawberry, sweet 2
66.Ethyl tetradecanoate179917962042 0.08 ± 0.00ether 1; waxy, soapy 2
67.Octadecane180018001800tr alkane
68.2-Hexadecanone1816180921120.06 ± 0.00 c0.06 ± 0.00 bc0.06 ± 0.00 bc0.05 ± 0.00 abctr afruit
69.Hexadecanal18281818 E2141 Etr atr abtr btr b
70.2-Phenylethyl benzoate18651859 E21890.15 ± 0.02 bc0.05 ± 0.01 a0.15 ± 0.01 bc0.15 ± 0.02 bc0.18 ± 0.01 cflower, honey 1; honey, rose 2
71.3-Heptadecanone18841880 E2155 E0.05 ± 0.00 a0.05 ± 0.00 a0.05 ± 0.01 a0.05 ± 0.01 a0.20 ± 0.01 b
72.n-Nonadecane1900190019000.20 ± 0.01 d0.17 ± 0.01 a0.19 ± 0.00 bcd0.19 ± 0.01 cd0.18 ± 0.00 abalkane
73.2-Heptadecanone19041900 E2245 Etr atr atr atr a
74.Methyl hexadecanoate192219212204 E0.18 ± 0.01 bc0.17 ± 0.01 bc0.19 ± 0.01 c0.17 ± 0.02 bc0.15 ± 0.00 a
75.Benzyl 4-methoxybenzoate19251922 G 0.11 ± 0.00 c0.09 ± 0.01 a0.10 ± 0.00 b0.10 ± 0.01 b0.09 ± 0.00 a
76.Hexadecanoic acid196719602931 E0.07 ± 0.01 btr a0.16 ± 0.02 c0.08 ± 0.02 b0.10 ± 0.01 b
77.Geranyl benzoate19821978 0.09 ± 0.00 c0.07 ± 0.00 a0.08 ± 0.00 bc0.08 ± 0.01 ab0.08 ± 0.00 bc
78.Ethyl hexadecanoate1997199322500.07 ± 0.00 bc0.08 ± 0.00 c0.04 ± 0.00 a0.06 ± 0.00 b1.31 ± 0.02 dwaxy
79.n-Eicosane2000200020000.05 ± 0.00 alkane
80.Octadecanal20362033 E24000.48 ± 0.01 c0.41 ± 0.03 a0.46 ± 0.00 b0.45 ± 0.03 b0.43 ± 0.01 aboil
81.n-Heneicosane2100210021005.50 ± 0.19 d4.16 ± 0.22 b4.69 ± 0.08 c4.49 ± 0.31 c3.71 ± 0.06 aalkane
82.Methyl oleate2105210324300.12 ± 0.00 a0.19 ± 0.01 b0.18 ± 0.01 b0.22 ± 0.01 d0.21 ± 0.00 cfat
83.Methyl linolenate21132108 E2590 E0.07 ± 0.00 b0.06 ± 0.00 a0.06 ± 0.00 a0.08 ± 0.00 c0.10 ± 0.00 d
84.Phytol21152115 E25710.05 ± 0.00 abtr atr a0.06 ± 0.01 b0.23 ± 0.00 cflower 1; balsamic, floral 2
85.(E)-Benzyl cinnamate21312134 E2769 Etr a tr b apricot, cherry, chocolate, floral, peach, pineapple 2
86.Linoleic acid21392134 E3168 E0.10 ± 0.01 c0.08 ± 0.01 b0.05 ± 0.00 a0.08 ± 0.00 bc1.53 ± 0.02 d
87.Oleic acid214721422430tr atr btr atr a0.07 ± 0.00 cfat
88.Ethyl 9-octadecenoate21622150 E2469 E 0.14 ± 0.00
89.n-Nonadecanol-121712156 E2637 Etr atr btr btr btr b
90.Ethyl linolenate21762169 E2621 Etr a0.12 ± 0.01 b0.15 ± 0.00 btr a1.56 ± 0.03 c
91.Hexadecanamide21892182 E2858 E0.14 ± 0.01 a0.18 ± 0.01 b0.20 ± 0.00 c0.22 ± 0.01 d0.32 ± 0.01 e
92.n-Butyl hexadecanoate21942188 E2419 Etr btr ctr a0.17 ± 0.01 d0.05 ± 0.01 c
93.1-Docosene21992189 0.10 ± 0.00 c0.09 ± 0.00 b0.10 ± 0.00 c0.10 ± 0.01 c0.07 ± 0.00 a
94.n-Docosane2200220022000.98 ± 0.05 d0.72 ± 0.04 ab0.83 ± 0.02 c0.79 ± 0.05 bc0.71 ± 0.02 aalkane
95.Tributyl acetylcitrate22582250 E tr a tr a
96.n-Tricosane23002300230011.36 ± 0.39 e8.54 ± 0.44 b9.94 ± 0.17 d9.14 ± 0.55 c7.57 ± 0.12 aalkane
97.2-Ethylhexyl p-methoxy cinnamate23492339 F3122 Ftr btr abtr btr btr b
98.4,8,12,16-Tetramethyl heptadecan-4-olide23642364 E 0.50 ± 0.01 d0.42 ± 0.02 c0.49 ± 0.01 d0.38 ± 0.02 b0.27 ± 0.00 a
99.(Z)-9-Octadecenamide23982397 E3265 E0.44 ± 0.04 a0.52 ± 0.02 b0.62 ± 0.01 c0.64 ± 0.03 c1.00 ± 0.06 d
100.n-Tetracosane2400240024000.27 ± 0.01 a0.37 ± 0.01 c0.43 ± 0.01 e0.40 ± 0.02 d0.35 ± 0.00 balkane
101.Heneicosanol24032402 E2995 E1.23 ± 0.04 c0.97 ± 0.05 a1.14 ± 0.02 b1.14 ± 0.06 b1.00 ± 0.02 a
102.n-Pentacosane2500250025006.13 ± 0.20 e4.80 ± 0.28 b5.63 ± 0.08 d5.11 ± 0.28 c4.18 ± 0.07 aalkane
103.n-Hexacosane2600260026000.43 ± 0.01 d0.36 ± 0.02 b0.40 ± 0.01 c0.37 ± 0.02 b0.32 ± 0.01 aalkane
104.Benzoic acid hexadecyl ester26652664 F 0.09 ± 0.00 a0.09 ± 0.01 a0.10 ± 0.00 a0.09 ± 0.02 a0.12 ± 0.01 b
105.n-Heptacosane27002700270015.40 ± 0.40 c13.93 ± 0.79 b14.66 ± 0.17 bc13.90 ± 1.15 b11.21 ± 0.26 aalkane
106.Benzyl oleate27572758 G 0.10 ± 0.02 btr a0.05 ± 0.01 a0.14 ± 0.03 c0.16 ± 0.02 c
107.Benzyl linoleate27672764 E 1.74 ± 0.02 a1.72 ± 0.08 ab1.85 ± 0.01 b2.09 ± 0.07 c2.17 ± 0.06 c
108.Benzyl linolenate27782775 E 0.30 ± 0.00 a0.27 ± 0.01 a0.26 ± 0.04 a0.39 ± 0.01 b0.53 ± 0.02 c
109.n-Octacosane2800280028000.14 ± 0.00 a0.39 ± 0.01 b0.41 ± 0.03 b0.59 ± 0.04 c0.54 ± 0.03 calkane
110.Squalene28352836 E2865 E0.10 ± 0.00 a0.14 ± 0.03 b0.15 ± 0.00 b0.35 ± 0.02 c0.88 ± 0.02 d
111.n-Nonacosane2900290029007.27 ± 0.11 c7.03 ± 0.31 c6.22 ± 0.08 b5.64 ± 0.20 a6.06 ± 0.18 balkane
112.1-Hexacosanol29122906 0.08 ± 0.01 a0.16 ± 0.01 c0.14 ± 0.01 bc0.15 ± 0.02 bc0.15 ± 0.04 bc
113.n-Triacontane3000300030000.17 ± 0.01 a0.32 ± 0.02 d0.24 ± 0.01 b0.23 ± 0.01 b0.27 ± 0.02 calkane
114.γ-Tocopherol30753074 tr abtr a0.05 ± 0.01 b0.12 ± 0.01 c
115.n-Untriacontane3100310031006.73 ± 0.20 c6.46 ± 0.38 b4.41 ± 0.13 a4.51 ± 0.08 a6.22 ± 0.24 balkane
116.Vitamin E (D-α-tocopherol)31543149 E 0.14 ± 0.00 a0.50 ± 0.03 c0.25 ± 0.01 b0.99 ± 0.01 d1.76 ± 0.11 e
Total compounds identified/representing % of total volatiles105/77.47100/74.12104/76.0793/74.1092/76.69
Grouped compounds (%)
Aromatics12.6712.5513.6414.1614.32
Aliphatic hydrocarbons (alkanes)54.9347.6548.4445.7241.47
Aromatic hydrocarbons0.040.040.040.18
Monoterpene hydrocarbons0.180.250.210.160.15
Oxygenated monoterpenes1.342.352.392.813.97
Oxygenated sesquiterpenes0.310.450.440.430.41
Alcohols and esters2.302.662.373.025.58
Aldehydes, ketones and lactones3.144.444.953.372.75
Acids1.271.711.751.443.12
Amides0.580.700.820.861.32
Diterpenoids0.050.040.050.060.23
Triterpenoids and tocopherols0.240.680.421.392.76
Phenylpropanoids and phenols0.120.160.140.140.40
Other (oxanes, heteroaromatics, etc.)0.350.480.460.420.44
# Compounds are listed in order of their elution from nonpolar BPX-5 capillary column. A Identified on the basis of GC-TOF/MS spectra based on comparison with NIST, MainLib, RepLib and Adams libraries and calculated KI with those reported in Adams and Nist, PubChem and ChemSpider databases. B Kováts retention indices calculated against C7-C30 n-alkanes on BPX-5 column. C Kováts retention indices on nonpolar DB-5 column reported in literature [27]. D Kováts retention indices calculated on polar carbowax 20 M column reported in literature (Flavornet, http://flavornet.org/flavornet.html) (accessed on 19 July 2021). E Kováts retention indices from the database http://webbook.nist.go (accessed on 19 July 2021). F Kováts retention indices from the database https://pubchem.ncbi.nlm.nih.gov (accessed on 19 July 2021). G Kováts retention indices from the database http://www.chemspider.com (accessed on 19 July 2021). 1 Odour description from http://flavornet.org. (accessed on 23 October 2021) (and without number indications) [25]; 2 Odour description from SAFC® Flavors & Fragrances (www.safcglobal.com) (accessed on 23 October 2021) [26]; tr—trace (≤ 0.04%); RSD%, average coefficient of variance of individual compounds. * n-Decane corresponds the percentage content of an alkane extracted initially from the plant material. Values within rows followed by the same superscript letter (a–e) do not differ statistically at p < 0.05 (Duncan test).
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Baranauskienė, R.; Venskutonis, P.R. Supercritical CO2 Extraction of Narcissus poeticus L. Flowers for the Isolation of Volatile Fragrance Compounds. Molecules 2022, 27, 353. https://doi.org/10.3390/molecules27020353

AMA Style

Baranauskienė R, Venskutonis PR. Supercritical CO2 Extraction of Narcissus poeticus L. Flowers for the Isolation of Volatile Fragrance Compounds. Molecules. 2022; 27(2):353. https://doi.org/10.3390/molecules27020353

Chicago/Turabian Style

Baranauskienė, Renata, and Petras Rimantas Venskutonis. 2022. "Supercritical CO2 Extraction of Narcissus poeticus L. Flowers for the Isolation of Volatile Fragrance Compounds" Molecules 27, no. 2: 353. https://doi.org/10.3390/molecules27020353

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