**Changes in the Qualitative and Quantitative Composition of Essential Oils of Clary Sage and Roman Chamomile During Steam Distillation in Pilot Plant Scale**

Susanne Wagner, Angela Pfleger, Michael Mandl and Herbert Böchzelt *Joanneum Research Forschungsgesellschaft mbH Graz, Resources, Institute of Water, Energy and Sustainability, Department for Plant Materials Sciences and Utilisation Austria* 

#### **1. Introduction**

140 Distillation – Advances from Modeling to Applications

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Clary sage (*Salvia sclarea* L.*)* from the genus Salvia is a biennial or short-living herbaceous perennial plant. Its leaves are united to a basal rosette in the first year, 12 to 25 cm long and 7 to 15 cm wide, ovate, cordate at the base, obtuse and long-stemmed. All the leaves are reticulate-rugose and hairy on both sides. The flowers seem loose to fairly dense, often branched paniculate. They reach approximately 2 cm and the large heart-shaped bracts are long, tapering and purple in early stages, later greenish white. Blossom: June or July to August. The seeds are 2 to 3 mm long nuts. Its native regions are the northern Mediterranean

Fig. 1. Clary sage

Changes in the Qualitative and Quantitative Composition of Essential Oils of

(1R, 5R)-1,5-epoxysalvial-4 (14)-ene, salvial-4(14)-en-1-one (Kintzios, 2000).

1996), (Ferichs & Rimbach , 1992).

**2.1 Cultivation and harvest** 

repeatability at each site.

**2. Methods** 

Unlike chamomile from the Roman chamomile only few pharmacological studies exist. The composition of Roman chamomile oil is very complex and so far app. 100 compounds were identified. Main compounds are the sesquiterpenes and sesquiterpene lactones from the germacranolide type (e.g. nobiline). It is characterized by the presence of terpenoids and saturated and unsaturated fatty acids with four or five carbon atoms, such as butyric, valeric, crotonic, angelic, tiglin and methacrylic acid. These are esterified with C3 to C6 alcohols such as n-butanol, isobutanol, isoamyl alcohol and 3 - methylpentane-1-ol. The relative proportion of total esters in the essential oil of Roman chamomile is known as the highest of all essential oil-producing plants. Furthermore, Roman chamomile flowers contain hydroperoxides (e.g. 1-β**-**hydroperoxyisonobiline, a sesquiterpene peroxide from germacran**e**-type and allylic hydroperoxides as well). The content of hydroperoxides in the dried drug varies and is decreased during prolonged storage (Barnes, et al., 2007), (Bajaj,

Both herbs were cultivated in the years 2002 and 2003 and Roman chamomile additionally in 2004 and 2010 by organic farming in three habitats (in 2010 only one habitat) of different altitude (Bad Blumau 285 m, Oberlungitz 400 m, St. Jakob 1000 m above sea level) in the East of Styria (Austria). Fields of the individual sites are divided into parcels a and b to ensure

The location of Bad Blumau (I) was located in the thermal spa area Blumau (Austria). Parcels were located within the spa recreation in order to attract interested visitors. The

Clary Sage and Roman Chamomile During Steam Distillation in Pilot Plant Scale 143

oxygenated isoprenoid compounds alcohols, ketones, esters and aldehydes) with various applications in medicine, pharmacology, perfumery, cosmetics, food preservation or as insect repellent. Essential oils also contain phenolic compounds and alkanes. They are usually produced by distillation, but also by mechanical processes from different plant parts, such as seeds, fruits, fruit peel, roots, leaves, needles and balsams. Unlike fatty oils essential oils are volatile. Several data published in literature show that the quality of essential oils and the oil yield depend on many different parameters e.g. plant development stage, genetic determination, plant organ, drying and storage conditions, soil structure, climate, insolation and weathering, habitat, harvesting methods, date of collection of plant material, analysis conditions used for identification of the compounds, etc. Relative amounts of the compounds and the chemical composition of essential oils may also vary due to the applied distillation process itself (lab or pilot plant scale) and the duration of the distillation. The essential oil of clary sage is a complex chemical mixture, consisting of up to 100 mostly terpenoid compounds. These are mainly (oxygenated) monoterpenes with small amounts of (oxygenated) sesquiterpenes. The main compounds are linalool (17.2%), linalyl acetate (14.3%), geraniol (6.5%), geranyl acetate (7.5%), terpineol (15.1%), nerol (5.5%), neryl acetate (5.2%) and sclareol (5.2%). Furthermore, α-pinene, β-pinene, camphene, myrcene, limonene, cis-and trans-ocimene, p-cymene, terpinolene, cis-3-hexen-1-ol, caryophyllene, terpinen-4-ol, citronellol, β-gurjunene, caryophyllene oxide, germacrene D, (2R, 5E)-2.12-epoxycaryophyll-5-ene, (2R, 5E)-caryophyllene-5-en-12-al, (2S, 5E)-caryophyllene-5-en-12-al, isospathulenol,

along with some other areas in Central Europe, North Africa and Central Asia. It is drought resistant and grows in dry, rocky places at an optimal soil pH of 4.5. Clary sage has a long history as medicinal herb and is currently grown mainly for the production of its essential oil reaching oil yields of 0.1 to 0.3%. (Board, 2003) (Dachler & Pelzmann, 1999) The clear, colourless oil is used in perfumes with a light, warm and sweet fragrance and as muscatel flavouring in food industry for vermouths, wines, and liqueurs. It is also used in aromatherapy for relieving anxiety, stress and fear, menstruation problems such as PMS (premenstrual syndrome) and cramping, and helping with insomnia, amenorrhea, dysmenorrhoea, pre-menopause, depression, fatigue, nervousness, migraine, varicose veins, haemorrhoids, oily skin and hair, spasmodic cough. It shows strong estrogenic, aphrodisiac and regenerative activity. (Lis-Balchin, 2006), (Kintzios, 2000) Sclareol, the essential oil main compound, is used in the perfume industry as a fixative and in the tobacco industry for flavouring. It is part of a large number of amber fragrances with woody note (Heinrich, et al., 2004), (Ferichs & Rimbach, 1992).

Fig. 2. Roman chamomile (Copyright Bernhard Bergmann)

Roman chamomile (*Chamaemelum nobile* L.) is a small (up to 80 cm), perennial plant found in dry fields and around gardens and cultivated grounds. It has daisy-like white flowers and is native to Western Europe, North America, and Argentina. Cultivation areas are mainly found in France (main producer), Belgium, Italy, Czech Republic, Slovakia, India, North America, Brazil and Argentina. Its stem is procumbent with ascending basic axis, the leaves are alternate, twice pinnate, finely dissected and fluffy to almost glabrous. The solitary, terminal flower heads rise eight to twelve inches above the ground and consist of prominent yellow disk flowers and silver-white ray flowers. Seeds are almost triangular, bald and shiny. (Dachler & Pelzmann, 1999) According to (Barnes, et al., 2007) Roman chamomile flower heads contain 0.4 – 1.75%. Blossom time is June and July, and its fragrance is sweet, crisp, fruity and herbaceous. The plant is used in tisanes, perfumes and cosmetics and to flavour foods. It is popular in aromatherapy, whose practitioners believe it to be a calming agent. The pharmacological profile of the Roman chamomile flower heads is similar to German chamomile (*Matricaria recutita* L.). It is used as mild sedative, anti-emetic and antispasmodic (Heinrich , et al., 2004), (Barnes, et al., 2007).

Essential oils are in general a complex mixture of natural compounds (terpene hydrocarbons like mono-, sesqui- and diterpenes - cyclic or non-cyclic – and their corresponding oxygenated isoprenoid compounds alcohols, ketones, esters and aldehydes) with various applications in medicine, pharmacology, perfumery, cosmetics, food preservation or as insect repellent. Essential oils also contain phenolic compounds and alkanes. They are usually produced by distillation, but also by mechanical processes from different plant parts, such as seeds, fruits, fruit peel, roots, leaves, needles and balsams. Unlike fatty oils essential oils are volatile. Several data published in literature show that the quality of essential oils and the oil yield depend on many different parameters e.g. plant development stage, genetic determination, plant organ, drying and storage conditions, soil structure, climate, insolation and weathering, habitat, harvesting methods, date of collection of plant material, analysis conditions used for identification of the compounds, etc. Relative amounts of the compounds and the chemical composition of essential oils may also vary due to the applied distillation process itself (lab or pilot plant scale) and the duration of the distillation.

The essential oil of clary sage is a complex chemical mixture, consisting of up to 100 mostly terpenoid compounds. These are mainly (oxygenated) monoterpenes with small amounts of (oxygenated) sesquiterpenes. The main compounds are linalool (17.2%), linalyl acetate (14.3%), geraniol (6.5%), geranyl acetate (7.5%), terpineol (15.1%), nerol (5.5%), neryl acetate (5.2%) and sclareol (5.2%). Furthermore, α-pinene, β-pinene, camphene, myrcene, limonene, cis-and trans-ocimene, p-cymene, terpinolene, cis-3-hexen-1-ol, caryophyllene, terpinen-4-ol, citronellol, β-gurjunene, caryophyllene oxide, germacrene D, (2R, 5E)-2.12-epoxycaryophyll-5-ene, (2R, 5E)-caryophyllene-5-en-12-al, (2S, 5E)-caryophyllene-5-en-12-al, isospathulenol, (1R, 5R)-1,5-epoxysalvial-4 (14)-ene, salvial-4(14)-en-1-one (Kintzios, 2000).

Unlike chamomile from the Roman chamomile only few pharmacological studies exist. The composition of Roman chamomile oil is very complex and so far app. 100 compounds were identified. Main compounds are the sesquiterpenes and sesquiterpene lactones from the germacranolide type (e.g. nobiline). It is characterized by the presence of terpenoids and saturated and unsaturated fatty acids with four or five carbon atoms, such as butyric, valeric, crotonic, angelic, tiglin and methacrylic acid. These are esterified with C3 to C6 alcohols such as n-butanol, isobutanol, isoamyl alcohol and 3 - methylpentane-1-ol. The relative proportion of total esters in the essential oil of Roman chamomile is known as the highest of all essential oil-producing plants. Furthermore, Roman chamomile flowers contain hydroperoxides (e.g. 1-β**-**hydroperoxyisonobiline, a sesquiterpene peroxide from germacran**e**-type and allylic hydroperoxides as well). The content of hydroperoxides in the dried drug varies and is decreased during prolonged storage (Barnes, et al., 2007), (Bajaj, 1996), (Ferichs & Rimbach , 1992).
