**1. Introduction**

The high biodiversity of Guatemala, caused by the great variety of microclimates and the convergence of the flora of North and South America, presents plants that have developed a large number of secondary metabolites to fulfill functions of defense and interaction with the environment. Many of these metabolites have biological and pharmacological activities that are used by communities, through the use of plants for the treatment of different diseases [1]. In this way, many investigations have been carried out aimed at determining the composition and biological activity of the metabolites of different medicinal plants used in Guatemala [2–5].

One of the biodiverse plants of Guatemala, which also grows in neighboring countries and for which no medicinal uses have been reported in Guatemala, is *Stevia serrata* Cav. [6] whose essential oil presents chamazulene in high proportions. Chamazulene is a substance of intense blue color of high economic value, which has been shown to have a high anti-inflammatory activity [7].

The genus *Stevia* belongs to the Asteraceae family within the Eupatorieae tribe [8]. It is a New World genus distributed from the south of the United States to Argentina and the highlands of Brazil, passing through Mexico, the Central American countries, and the South American Andes [9, 10]. The records indicate that the genus is not represented in the Antilles or the Amazon. The members of the *Stevia* genus are found mainly at altitudes between 500 and 3500 m. Although they usually grow in semidry mountainous terrain, their habitats range from meadows, leafy forests, forested mountain slopes, coniferous forests, to subalpine vegetation [8].

The genus *Stevia* consists of between 220 and 230 accepted species. Of these, only about 34 (15%) have some type of ethnobotanical record that relate uses with common names of the species. Of these 34 species, only the South American species *Stevia rebaudiana* (Bertoni) Bertoni presents records of outstanding use because its sweet leaves are used for imparting sweetness to beverages and foods [8, 12]. Due to this, *S. rebaudiana* is of great economic importance internationally, given its intensive commercialization due to its use as a natural low calorie sweetener [8].

The sesquiterpenoids are by far the majority and characteristic constituents of the aerial parts and roots of the *Stevia* genus. The overwhelming majority of these compounds belong to the guaiane, longipinane, and germacrene groups [8]. Derivatives of longipinene have been isolated and elucidated mainly in roots of *S. eupatoria*, *S. porphyria*, and *S. pilosa* in Mexico, in *S. triflora* from Venezuela, and in *S. lucida* of Colombia [13–18]. Diterpene glycosides have been isolated from commercial extracts of *S. rebaudiana* leaves in Malaysia [19, 20]. The composition of the essential oil of plants of the genus *Stevia* has been determined in leaves of *S. urticifolia* in Brazil being the main components found the oxygenated sesquiterpene α-cadinol (8.6%) and the sesquiterpene hydrocarbon germacrene D (10.4%) [21].

On the other hand, the composition of the essential oil of *S. rebaudiana* leaves analyzed in Nigeria showed carvacrol (67.89%), caryophyllene oxide (23.50%), spathulenol (15.41%), cardinol (5.59%), α-pinene (3.75%), ibuprofen (1.79%), isopinocarveol (1.26%), and α-caryophyllene (1.15%) as the main components found [22].

Other types of compounds isolated in plants of this genus include four flavonoids isolated from the aerial parts of *S. urticifolia* in Brazil [23], two triterpenes isolated from the roots of *S. viscida* and *S. eupatoria* from Mexico [24], the breviarolide and guaianolide isolated from the aerial parts of *S. breviaristata* from Argentina [25], and the stephalic acid isolated from the whole plant of *S. polycephala* from Mexico [26]. Nineteen hydroxycinnamic acid derivatives were successfully characterized in *S. rebaudiana* leaves: three monocaffeoylquinic acids (Mr354), seven dicaffeoylquinic acids (Mr516), one *p*-coumaroylquinic acid (Mr338), one feruloylquinic acid (Mr368), two caffeoyl-feruloylquinic acids (Mr530), three caffeoylshikimic acids (Mr336), and two tricaffeoylquinic acids (Mr678) [12].

Likewise, two new *stevia* amino acid sweeteners have been synthesized from natural stevioside: *stevia* glycine ethyl ester and *stevia* L-alanine methyl ester. The sweetness intensity rate of the new sweeteners was higher than sucrose, and they also had a clean sweetness without the unpleasant bitter aftertaste of stevioside [27]. *Stevia* products have been elaborated as an infusion with suitable organoleptic characteristics using a formulation of 80–85% of leaves + dried flowers of anise (*Tagetes filifolia* Lag.) and 15–20% of dried *stevia* leaves (*S. rebaudiana*) [28].

**205**

**Figure 1.**

*Chamazulene carboxylic acid.*

*Seed Propagation and Constituents of the Essential Oil of* Stevia serrata *Cav. from Guatemala*

As for the *S. serrata* plant, it is distributed from southern Arizona, New Mexico and Texas to northern Oaxaca, and from Chiapas to Honduras,

Colombia, Venezuela, and Ecuador. In Guatemala it is found in the departments of Chimaltenango, Guatemala, Huehuetenango, Quetzaltenango, El Quiché,

the corolla and alternating with 3–5 scales, 0.2–0.7 mm long [6, 11].

As for the chemistry of *S. serrata*, five new derivatives of longipinene have been isolated and elucidated from the roots of the plant in Mexico, these being 7β,9α-diangeloyloxy-8α-hydroxylongipinan-1-one; 8β,9α-diangeloyloxy-9αhydroxylongipinan-1-one; 7β,9α-diangeloyloxy-8α-acetyloxylongipinan-1-one; 7β,9α-diangeloyloxy-8α-acetyloxylongipin-2-en-1-one; and 7β-angeloyloxy-8αisobutyryloxylongipin-2-en-1-one [29]. Likewise, in Mexico, two new prochamazulene sesquiterpene lactones from the dried leaves of *S. serrata* from Mexico were isolated and identified: steviserrolide A and steviserrolide B [30]. The presence of the R enantiomer of chamazulene carboxylic acid (**Figure 1**) of *S. serrata* from

Regarding studies of the essential oil of the plant, the distillation of 178 g of flowers of *S. serrata* from Mexico provided 700 mg of the blue essential oil, which yielded 320 mg of chamazulene [32]. The compounds found in highest concentration in the essential oil of *S. serrata* from Guatemala were the sesquiterpenes chamazulene (60.1%), (*E*)-nerolidol (7.3%), caryophyllene oxide (6.3%), and germacrene D (5.4%) [33], which are shown in **Figure 2**. Chamazulene is produced from prochamazulenic sesquiterpenlactones. Among these precursors, matricine (**Figure 2**) and the carboxylic acid of chamazulene, among others, have been identified, which are present in the plant and are transformed into chamazulene by the action of the temperature during the steam extraction process [31]. Other compounds isolated from the plant include the methyl-ripariochromene A from the

The plant, known in Mexico as "tlachichinole," was used in decoction of the aerial parts for the washing of infected pimples [8], while the "donkey chili" or "sheep tail" is used as medicine to treat intestinal discomforts in Honduras [35]; the

The species grows along pastures and roadsides in various habitats from *Yucca*-*Opuntia* scrub, sand pine woods, steep rock outcrops in *Quercus*-*Acacia* grasslands, and pastured slopes, usually between 900 and 2800 m. The plants prefer sunny, stony, well-drained places but also grow in moist pastures and other flat areas [6, 11]. They grow as erect perennial herbs to 0.6–1 m, the stems single to many, puberulent to densely pilose. Leaves alternate, scattered and often crowded, sessile to subsessile, serrate toward the apex, 2.5–6.5 cm long, 0.2–1.5 cm wide, apex rounded to acute. Capitula 5–9 mm, phyllaries 3.5–6 mm long, 0.7–1 mm wide, puberulent with numerous glandular dots. Corollas white, 3–5 mm long, often gland-dotted, lobes 1–1.5 mm long, puberulent. Achenes are usually heteromorphic, 2.2–4.2 mm long, hispid. Pappus of the four adelphocarps with 3–5 awns equaling

*DOI: http://dx.doi.org/10.5772/intechopen.88221*

Sacatepéquez, and Sololá [6, 11].

Central America was determined [31].

dried leaves of *S. serrata* cultivated in Japan [34].

#### *Seed Propagation and Constituents of the Essential Oil of* Stevia serrata *Cav. from Guatemala DOI: http://dx.doi.org/10.5772/intechopen.88221*

As for the *S. serrata* plant, it is distributed from southern Arizona, New Mexico and Texas to northern Oaxaca, and from Chiapas to Honduras, Colombia, Venezuela, and Ecuador. In Guatemala it is found in the departments of Chimaltenango, Guatemala, Huehuetenango, Quetzaltenango, El Quiché, Sacatepéquez, and Sololá [6, 11].

The species grows along pastures and roadsides in various habitats from *Yucca*-*Opuntia* scrub, sand pine woods, steep rock outcrops in *Quercus*-*Acacia* grasslands, and pastured slopes, usually between 900 and 2800 m. The plants prefer sunny, stony, well-drained places but also grow in moist pastures and other flat areas [6, 11]. They grow as erect perennial herbs to 0.6–1 m, the stems single to many, puberulent to densely pilose. Leaves alternate, scattered and often crowded, sessile to subsessile, serrate toward the apex, 2.5–6.5 cm long, 0.2–1.5 cm wide, apex rounded to acute. Capitula 5–9 mm, phyllaries 3.5–6 mm long, 0.7–1 mm wide, puberulent with numerous glandular dots. Corollas white, 3–5 mm long, often gland-dotted, lobes 1–1.5 mm long, puberulent. Achenes are usually heteromorphic, 2.2–4.2 mm long, hispid. Pappus of the four adelphocarps with 3–5 awns equaling the corolla and alternating with 3–5 scales, 0.2–0.7 mm long [6, 11].

As for the chemistry of *S. serrata*, five new derivatives of longipinene have been isolated and elucidated from the roots of the plant in Mexico, these being 7β,9α-diangeloyloxy-8α-hydroxylongipinan-1-one; 8β,9α-diangeloyloxy-9αhydroxylongipinan-1-one; 7β,9α-diangeloyloxy-8α-acetyloxylongipinan-1-one; 7β,9α-diangeloyloxy-8α-acetyloxylongipin-2-en-1-one; and 7β-angeloyloxy-8αisobutyryloxylongipin-2-en-1-one [29]. Likewise, in Mexico, two new prochamazulene sesquiterpene lactones from the dried leaves of *S. serrata* from Mexico were isolated and identified: steviserrolide A and steviserrolide B [30]. The presence of the R enantiomer of chamazulene carboxylic acid (**Figure 1**) of *S. serrata* from Central America was determined [31].

Regarding studies of the essential oil of the plant, the distillation of 178 g of flowers of *S. serrata* from Mexico provided 700 mg of the blue essential oil, which yielded 320 mg of chamazulene [32]. The compounds found in highest concentration in the essential oil of *S. serrata* from Guatemala were the sesquiterpenes chamazulene (60.1%), (*E*)-nerolidol (7.3%), caryophyllene oxide (6.3%), and germacrene D (5.4%) [33], which are shown in **Figure 2**. Chamazulene is produced from prochamazulenic sesquiterpenlactones. Among these precursors, matricine (**Figure 2**) and the carboxylic acid of chamazulene, among others, have been identified, which are present in the plant and are transformed into chamazulene by the action of the temperature during the steam extraction process [31]. Other compounds isolated from the plant include the methyl-ripariochromene A from the dried leaves of *S. serrata* cultivated in Japan [34].

The plant, known in Mexico as "tlachichinole," was used in decoction of the aerial parts for the washing of infected pimples [8], while the "donkey chili" or "sheep tail" is used as medicine to treat intestinal discomforts in Honduras [35]; the

**Figure 1.** *Chamazulene carboxylic acid.*

*Essential Oils - Oils of Nature*

vegetation [8].

found [22].

One of the biodiverse plants of Guatemala, which also grows in neighboring countries and for which no medicinal uses have been reported in Guatemala, is *Stevia serrata* Cav. [6] whose essential oil presents chamazulene in high proportions. Chamazulene is a substance of intense blue color of high economic value, which has

The genus *Stevia* belongs to the Asteraceae family within the Eupatorieae tribe [8]. It is a New World genus distributed from the south of the United States to Argentina and the highlands of Brazil, passing through Mexico, the Central American countries, and the South American Andes [9, 10]. The records indicate that the genus is not represented in the Antilles or the Amazon. The members of the *Stevia* genus are found mainly at altitudes between 500 and 3500 m. Although they usually grow in semidry mountainous terrain, their habitats range from meadows, leafy forests, forested mountain slopes, coniferous forests, to subalpine

The genus *Stevia* consists of between 220 and 230 accepted species. Of these, only about 34 (15%) have some type of ethnobotanical record that relate uses with common names of the species. Of these 34 species, only the South American species *Stevia rebaudiana* (Bertoni) Bertoni presents records of outstanding use because its sweet leaves are used for imparting sweetness to beverages and foods [8, 12]. Due to this, *S. rebaudiana* is of great economic importance internationally, given its intensive commercialization due to its use as a natural low calorie sweetener [8]. The sesquiterpenoids are by far the majority and characteristic constituents of the aerial parts and roots of the *Stevia* genus. The overwhelming majority of these compounds belong to the guaiane, longipinane, and germacrene groups [8]. Derivatives of longipinene have been isolated and elucidated mainly in roots of *S. eupatoria*, *S. porphyria*, and *S. pilosa* in Mexico, in *S. triflora* from Venezuela, and in *S. lucida* of Colombia [13–18]. Diterpene glycosides have been isolated from commercial extracts of *S. rebaudiana* leaves in Malaysia [19, 20]. The composition of the essential oil of plants of the genus *Stevia* has been determined in leaves of *S. urticifolia* in Brazil being the main components found the oxygenated sesquiterpene α-cadinol (8.6%) and the

On the other hand, the composition of the essential oil of *S. rebaudiana* leaves analyzed in Nigeria showed carvacrol (67.89%), caryophyllene oxide (23.50%), spathulenol (15.41%), cardinol (5.59%), α-pinene (3.75%), ibuprofen (1.79%), isopinocarveol (1.26%), and α-caryophyllene (1.15%) as the main components

Other types of compounds isolated in plants of this genus include four flavonoids isolated from the aerial parts of *S. urticifolia* in Brazil [23], two triterpenes isolated from the roots of *S. viscida* and *S. eupatoria* from Mexico [24], the breviarolide and guaianolide isolated from the aerial parts of *S. breviaristata* from Argentina [25], and the stephalic acid isolated from the whole plant of *S. polycephala* from Mexico [26]. Nineteen hydroxycinnamic acid derivatives were successfully characterized in *S. rebaudiana* leaves: three monocaffeoylquinic acids (Mr354), seven dicaffeoylquinic acids (Mr516), one *p*-coumaroylquinic acid (Mr338), one feruloylquinic acid (Mr368), two caffeoyl-feruloylquinic acids (Mr530), three caffeoyl-

Likewise, two new *stevia* amino acid sweeteners have been synthesized from natural stevioside: *stevia* glycine ethyl ester and *stevia* L-alanine methyl ester. The sweetness intensity rate of the new sweeteners was higher than sucrose, and they also had a clean sweetness without the unpleasant bitter aftertaste of stevioside [27]. *Stevia* products have been elaborated as an infusion with suitable organoleptic characteristics using a formulation of 80–85% of leaves + dried flowers of anise (*Tagetes filifolia* Lag.) and 15–20% of dried *stevia* leaves (*S. rebaudiana*) [28].

shikimic acids (Mr336), and two tricaffeoylquinic acids (Mr678) [12].

been shown to have a high anti-inflammatory activity [7].

sesquiterpene hydrocarbon germacrene D (10.4%) [21].

**204**

**Figure 2.** *From left to right, structures of chamazulene, caryophyllene oxide, germacrene D, and matricine.*

decoction of the "October flower" is used by the midwives to accelerate the contractions of the parturients during childbirth [36]. Oral administration of *S. serrata* essential oil from Guatemala produced a marked antinociceptive activity in mice in the formalin test [33].

The purpose of the study was to determine the composition of the essential oil of aerial parts of *S. serrata* from different localities of the Guatemalan highlands, to evaluate the variability of the content of chamazulene. The capability of propagation of plants of *S. serrata* was also determined by a seed propagation trial. Finally, the composition of the essential oil of the roots of the propagated plants was determined to compare it with the composition of the oil extracted from aerial parts of the plant.
