# Chemical Diversity inLippia alba (Mill.) N. E. Brown Germplasm

Authors: Arie Fitzgerald Blank; Lídia Cristina Alves Camêlo; Maria de Fátima Arrigoni-Blank; José Baldin Pinheiro; Thiago Matos Andrade; Edenilson dos Santos Niculau; Péricles Barreto Alves
Journal: The Scientific World Journal
 (2015)

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## Abstract

The aim of this study was to perform chemical characterization ofLippia alba accessions from the Active Germplasm Bank of the Federal University of Sergipe. A randomized block experimental design with two replications was applied. The analysis of the chemical composition of the essential oils was conducted using a gas chromatograph coupled to a mass spectrometer. The chemical composition of the essential oils allowed the accessions to be allocated to the following six groups: group 1: linalool, 1,8-cineole, and caryophyllene oxide; group 2: linalool, geranial, neral, 1,8-cineol, and caryophyllene oxide; group 3: limonene, carvone, and sabinene; group 4: carvone, limonene, g-muurolene, and myrcene; group 5: neral, geranial, and caryophyllene oxide; and group 6: geranial, neral, o-cymene, limonene, and caryophyllene oxide.


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## Body

## 1. Introduction

The Verbenaceae family consists of approximately 175 genera and 2,800 species distributed in tropical and subtropical regions worldwide and in temperate regions of the southern hemisphere. In addition, a few species are found in temperate regions of the northern hemisphere [1]. The genusLippia includes several plant species of medicinal interest and comprises approximately 200 shrub species with a pantropical distribution and approximately 150 species distributed across rupestrian grasslands and tropical savannas (cerrados) in Brazil [2]. The speciesLippia alba (Mill.) N.E. Brown, also known asLippia geminata HBK andLantana alba (Mill), is a shrub with a height of approximately 3 meters [3]. In traditional Brazilian medicine, this species is popularly known as lemon balm [4].This species is considered by some authors [5] to be promising for use in the pharmaceutical, aromatic, and perfumery industries and may also be suitable for the agricultural chemical industry because of its proven antifungal, insecticidal, and repellent properties. The essential oil obtained fromL. alba has been recognized as a potential source of several commercially important terpenoid compounds [6].The chemical composition of essential oils consists of a mixture of many organic compounds in various concentrations, ranging from very low quantities (traces) to major compounds. Therefore, the variability of chemical types is a cause for concern from the standpoint of the use of essential oils as herbal medicines because some compounds may be unsuitable for achieving a desired result. This problem regarding medicinal plants is common in Brazil [7].Essential oils are primarily produced by the plant leaves and are formed through the secondary metabolism of plants. The typical compounds of this type include mono- and sesquiterpenes. Both the oil composition and plant yield, including biomass, are directly influenced by environmental factors, which represent a challenge for producers in establishing productive and stable genotypes and maintaining the chemical uniformity demanded by the industry [5].The rich pharmacological potential ofL. alba is related to the wide chemical variability of its essential oils. This variability allows the classification of this species into chemotypes, which can be defined according to the major chemical components of the essential oils [8]. The essential oils of some chemotypes identified fromL. alba differ in their chemical composition, with citral, carvone, and linalool representing the major components identified [9]. The limonene-carvone chemotype is characterized by the presence of limonene and carvone and the absence of neral and geranial (citral). Limonene is used as a solvent in cleaning products, foodstuffs, and the cosmetics industry. Carvone is used as a carminative and in cosmetic products and has bactericidal and fungicidal properties [10].The degree of variability in the active ingredient found in a medicinal species should be very low so that the drugs produced from that species are safe and effective [8]. Therefore, the identification and correct classification of chemotypes, which exhibit distinct active ingredients in medicinal plants, are of great importance for maintaining quality, planning cultivation, and obtaining phytochemicals that do not impair users’ health [11].Thus, the aim of this study was to perform chemical characterization of accessions ofL. alba (Mill.) NE Br. from the Active Germplasm Bank (AGB) of the Federal University of Sergipe.


## 2. Materials and Methods

### 2.1. Plant Materials

The experiment was conducted at the “Campus Rural da UFS” experimental farm, located in the municipality of São Cristóvão, state of Sergipe, at a latitude of 11°00′ S and longitude of 37°12′ W. Plants of 48 accessions ofL. alba from the AGB at UFS were evaluated (Table 1).Table 1
Identification and origin ofL. alba accessions from the Active Germplasm Bank at the Federal University of Sergipe.
Accession
Municipality/state
Origin
UFS herbarium code
LA-01
ABC-Distrito Federal
University of Brasília
14784
LA-02
Araguaína-Tocantins
University of Brasília
14785
LA-03
Atibaia-São Paulo
University of Brasília
13466
LA-04
Botucatu-São Paulo
University of Brasília
13501
LA-08
Brasília-Distrito Federal
University of Brasília
13475
LA-09
Brasília-Distrito Federal
University of Brasília
14786
LA-10
Brasília-Distrito Federal
University of Brasília
13495
LA-13
Fortaleza-Ceará
Federal University of Ceará
13488
LA-15
Florianópolis-Santa Catarina
University of Brasília
13486
LA-17
Brasília-Distrito Federal
University of Brasília
13494
LA-19
Brasília-Distrito Federal
University of Brasília
13491
LA-20
Ilhéus-Bahia
University of Brasília
14787
LA-21
Brasília-Distrito Federal
University of Brasília
13493
LA-22
Lavras-Minas Gerais
University of Brasília
13476
LA-24
Luziânia-Goiás
University of Brasília
13477
LA-27
Piracicaba-São Paulo
University of Brasília
13443
LA-28
Brasília-Distrito Federal
University of Brasília
13487
LA-29
Planaltina de Goiás-Goiás
University of Brasília
13485
LA-30
Posse-Goiás
University of Brasília
13454
LA-32
Rio de Janeiro-Rio de Janeiro
University of Brasília
13480
LA-36
Brasília-Distrito Federal
University of Brasília
13472
LA-37
Brasília-Distrito Federal
University of Brasília
13455
LA-39
Brasília-Distrito Federal
University of Brasília
13497
LA-40
Brasília-Distrito Federal
University of Brasília
13456
LA-41
Curitiba-Paraná
University of Brasília
13484
LA-42
Brasília-Distrito Federal
University of Brasília
13444
LA-43
Brasília-Distrito Federal
University of Brasília
13490
LA-44
Brasília-Distrito Federal
University of Brasília
14788
LA-45
Viçosa-Minas Gerais
University of Brasília
13498
LA-49
Aracaju-Sergipe
Federal University of Sergipe
13471
LA-52
Rio Real-Bahia
Federal University of Sergipe
13481
LA-53
Telha-Sergipe
Federal University of Sergipe
13446
LA-54
Rio Real-Bahia
Federal University of Sergipe
13478
LA-55
Rio Real-Bahia
Federal University of Sergipe
13468
LA-56
Rio Real-Bahia
Federal University of Sergipe
13465
LA-57
Rio Real-Bahia
Federal University of Sergipe
13469
LA-58
Rio Real-Bahia
Federal University of Sergipe
13482
LA-59
Rio Real-Bahia
Federal University of Sergipe
13500
LA-60
Rio Real-Bahia
Federal University of Sergipe
13499
LA-61
Rio Real-Bahia
Federal University of Sergipe
13479
LA-62
Rio Real-Bahia
Federal University of Sergipe
13451
LA-63
Santana do São Francisco-Sergipe
Federal University of Sergipe
13445
LA-67
Santana do São Francisco-Sergipe
Federal University of Sergipe
13464
LA-68
Santana do São Francisco-Sergipe
Federal University of Sergipe
14789
LA-69
Gararu-Sergipe
Federal University of Sergipe
13467
LA-70
Cristinápolis-Sergipe
Federal University of Sergipe
13473
LA-71
Paripiranga-Sergipe
Federal University of Sergipe
13447
LA-72
Traipú-Alagoas
Federal University of Sergipe
13496A randomized block experimental design with two replications was employed. Each plot consisted of three plants. Spacing of 1.5 meters was maintained both between individual plants and between rows. The fertilization applied in the field was 5 kg of cattle manure per pit. Culture practices such as weeding and hydration were performed whenever necessary.


### 2.2. Distillation and Analysis of Essential Oils

The plants were cut at a height of 30 cm from the soil, and the leaves were dried in an incubator with forced airflow at a temperature of 40°C for five days [12]. After drying, the leaves were weighed on an electronic scale, and essential oils were extracted using the hydrodistillation method in a Clevenger apparatus. For hydrodistillation, 75 g of dry leaves and 2.0 L of distilled water were used per flask, and the distillation period was 120 minutes after the initiation of water vapor condensation in the Clevenger apparatus. After extraction, the essential oils were collected and stored in a freezer in amber glass vials.Chemical analysis of the essential oil was performed at the Laboratory of Chromatography of the Department of Chemistry at the Federal University of Sergipe.Qualitative analysis of the chemical composition of the essential oil was performed in a gas chromatograph coupled to a mass spectrometer (GC-MS, model QP 5050A, Shimadzu) equipped with an AOC-20i autosampler (Shimadzu) and a fused-silica capillary column (5% phenyl, 95% dimethylpolysiloxane, 30 m × 0.25 mm i.d., and film thickness of 0.25μm, J&W Scientific) using helium as the carrier gas at a flow rate of 1.2 mL min−1. The temperature ramp was 50°C for 2 min, followed by an increase of 4°C min−1 until reaching 200°C, then an increase of 15°C until reaching 300°C, after which a constant temperature was maintained for 15 min. The injector temperature was maintained at 250°C, and that of the detector (or interface) was maintained at 280°C. A volume of 0.5 μL was injected using ethyl acetate. The partition rate of the injected volume was 1 : 100, and the column pressure was 64.20 kPa. The MS conditions included an ion capture detector operated through electron impact and an impact energy of 70 eV, a scan rate of 1,000, a scan interval of 0.50 fragment/s, and a fragment mass range between 40 Da and 500 Da.Quantitative analysis of the chemical constituents was performed by flame ionization gas chromatography (FID), using a Shimadzu GC-17A (Shimadzu Corporation, Kyoto, Japan) instrument, under the following operational conditions: capillary ZB-5MS column (5% phenyl-arylene-95%-dimethylpolysiloxane) fused-silica capillary column (30 m × 0.25 mm i.d. × 0.25μm film thickness) from Phenomenex (Torrance, CA, USA), under the same conditions as reported for the GC-MS. Quantification of each constituent was estimated by area normalization (%). Compound concentrations were calculated from the GC peak areas and they were arranged in order of GC elution.The essential oil components were identified by comparing their mass spectra with the available spectra in the equipment database (NIST05 and WILEY8). Additionally, the measured retention indices were compared with those in the literature [13]. The relative retention indices (RRI) were determined using the van den Dool and Kratz [14] equation and a homologous series of n-alkanes (C8–C18) injected under the chromatography conditions described above.


### 2.3. Statistical Analysis

The data were subjected to variance analysis, and the means were compared using the Scott-Knott test (p≤0.05). The chemical composition data were analyzed through two multivariate analysis methods: principal component analysis (PCA) and arrangement analysis (cluster) based on the similarity and distribution of the compounds, using Statistica software, version 7.0.


## 2.1. Plant Materials

The experiment was conducted at the “Campus Rural da UFS” experimental farm, located in the municipality of São Cristóvão, state of Sergipe, at a latitude of 11°00′ S and longitude of 37°12′ W. Plants of 48 accessions ofL. alba from the AGB at UFS were evaluated (Table 1).Table 1
Identification and origin ofL. alba accessions from the Active Germplasm Bank at the Federal University of Sergipe.
Accession
Municipality/state
Origin
UFS herbarium code
LA-01
ABC-Distrito Federal
University of Brasília
14784
LA-02
Araguaína-Tocantins
University of Brasília
14785
LA-03
Atibaia-São Paulo
University of Brasília
13466
LA-04
Botucatu-São Paulo
University of Brasília
13501
LA-08
Brasília-Distrito Federal
University of Brasília
13475
LA-09
Brasília-Distrito Federal
University of Brasília
14786
LA-10
Brasília-Distrito Federal
University of Brasília
13495
LA-13
Fortaleza-Ceará
Federal University of Ceará
13488
LA-15
Florianópolis-Santa Catarina
University of Brasília
13486
LA-17
Brasília-Distrito Federal
University of Brasília
13494
LA-19
Brasília-Distrito Federal
University of Brasília
13491
LA-20
Ilhéus-Bahia
University of Brasília
14787
LA-21
Brasília-Distrito Federal
University of Brasília
13493
LA-22
Lavras-Minas Gerais
University of Brasília
13476
LA-24
Luziânia-Goiás
University of Brasília
13477
LA-27
Piracicaba-São Paulo
University of Brasília
13443
LA-28
Brasília-Distrito Federal
University of Brasília
13487
LA-29
Planaltina de Goiás-Goiás
University of Brasília
13485
LA-30
Posse-Goiás
University of Brasília
13454
LA-32
Rio de Janeiro-Rio de Janeiro
University of Brasília
13480
LA-36
Brasília-Distrito Federal
University of Brasília
13472
LA-37
Brasília-Distrito Federal
University of Brasília
13455
LA-39
Brasília-Distrito Federal
University of Brasília
13497
LA-40
Brasília-Distrito Federal
University of Brasília
13456
LA-41
Curitiba-Paraná
University of Brasília
13484
LA-42
Brasília-Distrito Federal
University of Brasília
13444
LA-43
Brasília-Distrito Federal
University of Brasília
13490
LA-44
Brasília-Distrito Federal
University of Brasília
14788
LA-45
Viçosa-Minas Gerais
University of Brasília
13498
LA-49
Aracaju-Sergipe
Federal University of Sergipe
13471
LA-52
Rio Real-Bahia
Federal University of Sergipe
13481
LA-53
Telha-Sergipe
Federal University of Sergipe
13446
LA-54
Rio Real-Bahia
Federal University of Sergipe
13478
LA-55
Rio Real-Bahia
Federal University of Sergipe
13468
LA-56
Rio Real-Bahia
Federal University of Sergipe
13465
LA-57
Rio Real-Bahia
Federal University of Sergipe
13469
LA-58
Rio Real-Bahia
Federal University of Sergipe
13482
LA-59
Rio Real-Bahia
Federal University of Sergipe
13500
LA-60
Rio Real-Bahia
Federal University of Sergipe
13499
LA-61
Rio Real-Bahia
Federal University of Sergipe
13479
LA-62
Rio Real-Bahia
Federal University of Sergipe
13451
LA-63
Santana do São Francisco-Sergipe
Federal University of Sergipe
13445
LA-67
Santana do São Francisco-Sergipe
Federal University of Sergipe
13464
LA-68
Santana do São Francisco-Sergipe
Federal University of Sergipe
14789
LA-69
Gararu-Sergipe
Federal University of Sergipe
13467
LA-70
Cristinápolis-Sergipe
Federal University of Sergipe
13473
LA-71
Paripiranga-Sergipe
Federal University of Sergipe
13447
LA-72
Traipú-Alagoas
Federal University of Sergipe
13496A randomized block experimental design with two replications was employed. Each plot consisted of three plants. Spacing of 1.5 meters was maintained both between individual plants and between rows. The fertilization applied in the field was 5 kg of cattle manure per pit. Culture practices such as weeding and hydration were performed whenever necessary.


## 2.2. Distillation and Analysis of Essential Oils

The plants were cut at a height of 30 cm from the soil, and the leaves were dried in an incubator with forced airflow at a temperature of 40°C for five days [12]. After drying, the leaves were weighed on an electronic scale, and essential oils were extracted using the hydrodistillation method in a Clevenger apparatus. For hydrodistillation, 75 g of dry leaves and 2.0 L of distilled water were used per flask, and the distillation period was 120 minutes after the initiation of water vapor condensation in the Clevenger apparatus. After extraction, the essential oils were collected and stored in a freezer in amber glass vials.Chemical analysis of the essential oil was performed at the Laboratory of Chromatography of the Department of Chemistry at the Federal University of Sergipe.Qualitative analysis of the chemical composition of the essential oil was performed in a gas chromatograph coupled to a mass spectrometer (GC-MS, model QP 5050A, Shimadzu) equipped with an AOC-20i autosampler (Shimadzu) and a fused-silica capillary column (5% phenyl, 95% dimethylpolysiloxane, 30 m × 0.25 mm i.d., and film thickness of 0.25μm, J&W Scientific) using helium as the carrier gas at a flow rate of 1.2 mL min−1. The temperature ramp was 50°C for 2 min, followed by an increase of 4°C min−1 until reaching 200°C, then an increase of 15°C until reaching 300°C, after which a constant temperature was maintained for 15 min. The injector temperature was maintained at 250°C, and that of the detector (or interface) was maintained at 280°C. A volume of 0.5 μL was injected using ethyl acetate. The partition rate of the injected volume was 1 : 100, and the column pressure was 64.20 kPa. The MS conditions included an ion capture detector operated through electron impact and an impact energy of 70 eV, a scan rate of 1,000, a scan interval of 0.50 fragment/s, and a fragment mass range between 40 Da and 500 Da.Quantitative analysis of the chemical constituents was performed by flame ionization gas chromatography (FID), using a Shimadzu GC-17A (Shimadzu Corporation, Kyoto, Japan) instrument, under the following operational conditions: capillary ZB-5MS column (5% phenyl-arylene-95%-dimethylpolysiloxane) fused-silica capillary column (30 m × 0.25 mm i.d. × 0.25μm film thickness) from Phenomenex (Torrance, CA, USA), under the same conditions as reported for the GC-MS. Quantification of each constituent was estimated by area normalization (%). Compound concentrations were calculated from the GC peak areas and they were arranged in order of GC elution.The essential oil components were identified by comparing their mass spectra with the available spectra in the equipment database (NIST05 and WILEY8). Additionally, the measured retention indices were compared with those in the literature [13]. The relative retention indices (RRI) were determined using the van den Dool and Kratz [14] equation and a homologous series of n-alkanes (C8–C18) injected under the chromatography conditions described above.


## 2.3. Statistical Analysis

The data were subjected to variance analysis, and the means were compared using the Scott-Knott test (p≤0.05). The chemical composition data were analyzed through two multivariate analysis methods: principal component analysis (PCA) and arrangement analysis (cluster) based on the similarity and distribution of the compounds, using Statistica software, version 7.0.


## 3. Results and Discussion

Among the compounds present in the essential oils from 48 accessions, 33 were identified and are listed according to their order of elution (Table2).Table 2
Chemical composition of the essential oil ofL. alba accessions from the Active Germplasm Bank at the Federal University of Sergipe.
Compound
RRI
Accession
LA-01
LA-02
LA-03
LA-04
LA-08
LA-09
LA-10
LA-13
α-thujene
924
0.00b
0.00b
0.43b
0.27b
0.00b
0.00b
0.00b
0.00b
Sabinene
969
0.00c
0.00c
0.00c
0.15c
0.00c
0.64b
0.00c
4.39a
1-Octen-3-ol
974
0.00b
0.00b
0.00b
0.28b
0.72a
0.00b
0.00b
0.00b
5-Methyl-6-hepten-2-one
981
0.00a
0.00a
0.46a
0.41a
0.34a
0.00a
0.00a
0.00a
Myrcene
988
0.00c
0.76c
2.78b
2.85b
0.31c
0.00c
0.00c
0.00c
o-Cymene
1022
0.00d
0.00d
0.11d
0.32d
0.00d
0.00d
0.00d
0.00d
Limonene
1024
0.00e
0.00e
10.24c
0.17e
0.00e
0.00e
0.00e
45.35a
1,8-Cineol
1026
7.22b
0.00d
0.00d
0.00d
0.00d
9.22a
0.00d
0.00d
γ-terpinene
1054
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
cis-sabinene hydrate
1065
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
cis-linalool oxide
1067
0.86b
0.00c
0.00c
0.00c
0.00c
1.20a
0.00c
0.00c
trans-linalool oxide
1084
0.65b
0.00d
0.00d
0.00d
0.00d
0.98a
0.00d
0.00d
Linalool
1095
84.73a
0.84f
0.89f
1.09f
0.95f
80.63b
1.32f
0.00f
Perylene
1102
0.00b
0.00b
0.10a
0.19a
0.00b
0.00b
0.00b
0.00b
(E)-isocitral
1177
0.00b
0.00b
0.50a
0.22b
0.00b
0.00b
0.00b
0.00b
Myrtenal
1195
0.67b
0.00c
0.29c
0.54b
0.56b
0.71b
0.00c
0.00c
Neral
1235
0.00f
32.14a
29.78c
30.28c
31.09c
0.00f
29.32c
0.00f
Carvone
1239
0.00e
0.00e
3.91d
0.33e
0.43e
0.00e
0.00e
39.58c
Geraniol
1249
0.00c
0.00c
0.71c
0.00c
0.00c
0.00c
0.00c
0.00c
Geranial
1264
0.00f
54.05a
47.66a
49.76a
49.45a
0.00f
48.06a
0.00f
Myrtanyl acetate
1324
0.00d
0.00d
0.13c
0.00d
0.24c
0.00d
0.00d
0.00d
Geranyl acetate
1379
0.00c
0.00c
0.35b
0.29b
0.00c
0.00c
0.82a
0.00c
β-elemene
1389
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.49d
0.00d
β-caryophyllene
1417
1.04c
0.42c
0.00c
0.78c
0.94c
1.11c
0.62c
0.00c
α-guaiene
1437
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-muurolene
1478
0.62c
0.00c
0.29c
0.00c
0.00c
0.52c
0.00c
0.00c
trans-calamenene
1521
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Elemol
1548
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Germacrene B
1559
0.57b
0.00c
0.00c
0.00c
0.00c
0.28c
0.00c
0.00c
(E)-nerolidol
1561
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Spathulenol
1577
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Caryophyllene oxide
1582
2.93d
11.33c
0.00e
10.69c
14.24c
3.66d
17.77b
0.00e
Humulene epoxide II
1608
0.00d
0.00d
0.15d
0.00d
0.00d
0.00d
0.59b
0.00d
Essential oil content (%)
2.53a
1.30d
2.06c
1.02d
0.79e
2.54a
0.86e
0.80e
LA-15
LA-17
LA-19
LA-20
LA-21
LA-22
LA-24
LA-27
α-thujene
924
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Sabinene
969
0.00c
0.00c
0.00c
0.71b
0.00c
0.29c
0.00c
0.64b
1-Octen-3-ol
974
0.42a
0.78a
0.77a
0.00b
0.00b
0.00b
0.00b
0.00b
5-Methyl-6-hepten-2-one
981
1.03a
0.91a
0.58a
0.00a
0.38a
0.00a
0.00a
0.00a
Myrcene
988
3.07b
0.67c
0.45c
0.76c
0.84c
0.00c
0.00c
0.21c
o-Cymene
1022
0.00d
0.00d
0.00d
1.07d
0.00d
0.00d
0.62d
0.00d
Limonene
1024
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
1,8-Cineol
1026
0.00d
0.00d
0.00d
6.62b
0.93d
9.17a
2.38c
7.83b
γ-terpinene
1054
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
cis-sabinene hydrate
1065
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
cis-linalool oxide
1067
0.00c
0.00c
0.00c
0.64b
0.00c
1.00a
0.00c
0.80b
trans-linalool oxide
1084
0.00d
0.00d
0.00d
0.00d
0.00d
0.23c
0.00d
0.61b
Linalool
1095
0.83f
1.43f
0.83f
57.69d
7.27f
84.45a
49.38e
75.79c
Perylene
1102
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
(E)-isocitral
1177
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Myrtenal
1195
0.00c
0.00c
0.95a
0.00c
0.34c
0.00c
0.00c
0.66b
Neral
1235
31.98a
28.91c
31.22c
11.51e
28.98c
0.00f
16.59e
0.58f
Carvone
1239
0.00e
0.00e
0.00e
0.00e
0.43e
0.00e
0.00e
0.00e
Geraniol
1249
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
Geranial
1264
50.54a
48.68a
49.09a
16.99e
44.91a
0.00f
26.18e
0.83f
Myrtanyl acetate
1324
0.00d
0.00d
0.53b
0.00d
0.00d
0.00d
0.00d
0.00d
Geranyl acetate
1379
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
β-elemene
1389
0.00d
0.25d
0.00d
0.00d
0.00d
0.00d
0.00d
0.46d
β-caryophyllene
1417
1.75b
1.03c
0.89c
0.83c
0.71c
0.29c
0.74c
1.59b
α-guaiene
1437
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-muurolene
1478
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
1.60b
trans-calamenene
1521
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Elemol
1548
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Germacrene B
1559
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
1.26a
(E)-nerolidol
1561
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Spathulenol
1577
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Caryophyllene oxide
1582
10.35c
16.86b
14.04c
3.18d
14.63c
4.35d
4.11d
0.00e
Humulene epoxide II
1608
0.00d
0.00d
0.00d
0.00d
0.29c
0.20c
0.00d
0.00d
Essential oil content (%)
1.09d
1.04d
0.89e
1.33d
0.76e
2.53a
2.50a
2.26b
LA-28
LA-29
LA-30
LA-32
LA-36
LA-37
LA-39
LA-40
α-thujene
924
0.00b
0.00b
0.00b
0.91a
0.00b
0.00b
0.00b
0.22b
Sabinene
969
0.00c
0.00c
0.27c
0.00c
0.24c
0.00c
0.53b
0.27c
1-Octen-3-ol
974
0.63a
0.00b
0.00b
0.31b
0.00b
1.19a
0.00b
0.63a
5-Methyl-6-hepten-2-one
981
0.00a
0.00a
0.72a
0.23a
0.46a
0.00a
0.35a
0.52a
Myrcene
988
0.00c
7.09a
0.40c
3.67b
0.17c
0.00c
0.20c
1.16c
o-Cymene
1022
0.00d
0.00d
3.71d
12.36a
3.96c
0.00d
5.30c
0.00d
Limonene
1024
0.00e
0.00e
7.08c
0.00e
6.80c
0.00e
8.55c
0.00e
1,8-Cineol
1026
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-terpinene
1054
0.00c
0.00c
1.05a
0.00c
1.03a
0.00c
1.01a
0.00c
cis-sabinene hydrate
1065
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.29b
cis-linalool oxide
1067
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.30c
trans-linalool oxide
1084
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Linalool
1095
0.91f
0.00f
0.92f
0.25f
0.85f
0.00f
0.92f
2.90f
Perylene
1102
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
(E)-isocitral
1177
0.00b
0.00b
0.00b
0.00b
0.34a
0.00b
0.57a
0.00b
Myrtenal
1195
0.72b
0.67b
0.00c
0.49b
0.00c
1.19a
0.00c
0.00c
Neral
1235
31.99a
28.09c
32.93a
21.94c
30.87c
23.76c
31.66a
29.95c
Carvone
1239
0.00e
0.56e
0.93e
0.39e
0.99e
1.16e
1.05e
0.00e
Geraniol
1249
0.00c
0.00c
0.00c
2.03b
0.00c
0.00c
0.00c
0.00c
Geranial
1264
49.73a
40.24c
49.07a
32.90c
51.32a
39.89c
48.00a
46.80a
Myrtanyl acetate
1324
0.00d
0.00d
0.00d
0.00d
0.00d
0.72a
0.00d
0.00d
Geranyl acetate
1379
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
β-elemene
1389
0.00d
1.19b
0.00d
0.00d
0.00d
0.00d
0.00d
1.88a
β-caryophyllene
1417
0.55c
0.00c
0.00c
3.69a
0.00c
1.54b
0.00c
1.31b
α-guaiene
1437
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-muurolene
1478
0.00c
0.00c
0.41c
0.00c
0.40c
0.00c
0.00c
0.00c
trans-calamenene
1521
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Elemol
1548
0.00d
0.00d
1.14c
0.00d
2.16a
0.00d
1.83a
0.00d
Germacrene B
1559
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
(E)-nerolidol
1561
0.00e
0.00e
1.14d
0.00e
2.16a
0.00e
1.83b
0.00e
Spathulenol
1577
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Caryophyllene oxide
1582
14.86c
10.89c
1.32e
20.46b
0.00e
28.09a
0.00e
11.43c
Humulene epoxide II
1608
0.00d
0.00d
0.00d
0.35c
0.00d
0.00d
0.00d
0.00d
Essential oil content (%)
0.58e
0.66e
1.06d
1.26d
2.21b
0.80e
1.84c
0.53e
LA-41
LA-42
LA-43
LA-44
LA-45
LA-49
LA-52
LA-53
α-thujene
924
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Sabinene
969
0.14c
0.28c
0.31c
0.00c
0.98b
0.57b
0.54b
0.17c
1-Octen-3-ol
974
0.63a
0.44a
0.00b
0.58a
0.00b
0.21b
0.00b
0.59a
5-Methyl-6-hepten-2-one
981
0.74a
0.00a
0.43a
0.51a
0.30a
0.26a
0.30a
0.34a
Myrcene
988
1.47c
2.04c
0.71c
0.42c
8.47a
0.00c
0.21c
0.00c
o-Cymene
1022
0.31d
0.00d
0.00d
0.00d
0.32d
11.50a
9.44b
4.49c
Limonene
1024
0.00e
0.00e
7.46c
0.00e
0.00e
0.00e
3.86d
0.00e
1,8-Cineol
1026
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-terpinene
1054
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.98a
0.00c
cis-sabinene hydrate
1065
0.00c
0.00c
0.00c
0.00c
0.46b
0.00c
0.00c
0.33b
cis-linalool oxide
1067
0.15c
0.00c
0.00c
0.00c
0.00c
0.27c
0.00c
0.00c
trans-linalool oxide
1084
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Linalool
1095
0.85f
2.22f
0.74f
0.73f
1.33f
0.33f
0.00f
0.57f
Perylene
1102
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
(E)-isocitral
1177
0.00b
0.00b
0.75a
0.00b
0.38a
0.00b
0.46a
0.00b
Myrtenal
1195
0.00c
0.00c
0.00c
0.68b
0.00c
0.00c
0.00c
0.00c
Neral
1235
30.88c
31.33c
34.86a
32.27a
28.57c
23.69c
29.97c
31.57a
Carvone
1239
0.00e
0.00e
3.56d
0.00e
0.00e
0.00e
0.00e
0.00e
Geraniol
1249
0.00c
0.00c
0.00c
0.00c
3.38a
0.00c
0.00c
0.00c
Geranial
1264
48.69a
48.59a
51.18a
49.88a
46.36a
39.39c
46.22a
48.37a
Myrtanyl acetate
1324
0.00d
0.00d
0.00d
0.50b
0.00d
0.00d
0.00d
0.00d
Geranyl acetate
1379
0.27b
0.00c
0.00c
0.00c
0.40b
1.03a
0.00c
0.00c
β-elemene
1389
0.79c
1.75a
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
β-caryophyllene
1417
0.88c
1.23c
0.00c
0.00c
2.84a
1.15c
1.60b
0.00c
α-guaiene
1437
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
1.66b
γ-muurolene
1478
0.00c
0.00c
0.00c
0.00c
0.33c
0.38c
0.00c
0.00c
trans-calamenene
1521
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Elemol
1548
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Germacrene B
1559
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
(E)-nerolidol
1561
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Spathulenol
1577
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
3.21a
Caryophyllene oxide
1582
12.46c
11.66c
0.00e
13.43c
5.37d
19.89b
6.05d
5.05d
Humulene epoxide II
1608
0.00d
0.00d
0.00d
0.00d
0.00d
0.66b
0.34c
1.45a
Essential oil content (%)
0.85e
0.90e
1.80c
0.69e
1.06d
1.18d
2.00c
0.76e
LA-54
LA-55
LA-56
LA-57
LA-58
LA-59
LA-60
LA-61
α-thujene
924
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Sabinene
969
0.59b
0.53b
0.00c
0.00c
0.00c
0.25c
0.00c
0.00c
1-Octen-3-ol
974
0.00b
0.00b
0.00b
0.00b
0.67a
0.52a
0.00b
0.00b
5-Methyl-6-hepten-2-one
981
0.00a
0.00a
0.00a
0.00a
0.65a
0.92a
0.00a
0.53a
Myrcene
988
0.00c
0.00c
0.17c
0.15c
1.27c
2.80b
0.00c
1.31c
o-Cymene
1022
9.55b
8.29b
0.00d
0.00d
2.93c
3.77c
6.47b
3.53c
Limonene
1024
3.97d
3.69d
18.48b
19.35b
0.00e
0.00e
2.60e
0.00e
1,8-Cineol
1026
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-terpinene
1054
0.90a
0.93a
0.00c
0.00c
0.00c
0.00c
0.85a
0.00c
cis-sabinene hydrate
1065
0.00c
0.00c
0.00c
0.00c
0.00c
0.22c
0.00c
0.00c
cis-linalool oxide
1067
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
trans-linalool oxide
1084
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Linalool
1095
0.00f
0.00f
0.32f
0.19f
2.83f
2.31f
0.00f
2.30f
Perylene
1102
0.00b
0.00b
0.00b
0.00b
0.14a
0.13a
0.00b
0.00b
(E)-isocitral
1177
0.85a
0.44a
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Myrtenal
1195
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
Neral
1235
29.99c
30.98c
0.00f
0.00f
33.20a
33.93a
33.46a
35.31a
Carvone
1239
0.00e
0.00e
77.74a
77.20a
0.00e
0.00e
0.00e
0.00e
Geraniol
1249
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
Geranial
1264
44.02a
47.27a
0.00f
0.00f
51.42a
51.35a
48.61a
52.86a
Myrtanyl acetate
1324
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Geranyl acetate
1379
0.00c
0.00c
0.00c
0.00c
1.01a
0.83a
0.00c
0.83a
β-elemene
1389
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
β-caryophyllene
1417
2.18b
1.88b
0.00c
0.00c
0.00c
0.00c
3.16a
0.00c
α-guaiene
1437
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-muurolene
1478
0.00c
0.00c
1.99b
1.95b
0.00c
0.00c
0.00c
0.00c
trans-calamenene
1521
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
Elemol
1548
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Germacrene B
1559
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
(E)-nerolidol
1561
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Spathulenol
1577
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
0.00e
Caryophyllene oxide
1582
7.54d
5.96d
0.00e
0.00e
5.00d
2.59d
4.85d
3.00d
Humulene epoxide II
1608
0.38c
0.00d
0.00d
0.00d
0.87b
0.36c
0.00d
0.31c
Essential oil content (%)
1.93c
2.33b
1.60c
2.66a
0.66e
1.00d
1.73c
0.66e
LA-62
LA-63
LA-67
LA-68
LA-69
LA-70
LA-71
LA-72
α-thujene
924
0.00b
0.25b
0.00b
0.00b
0.00b
0.00b
0.24b
0.00b
Sabinene
969
0.00c
0.75b
0.00c
0.81b
0.37b
0.00c
0.55b
0.51b
1-Octen-3-ol
974
0.00b
0.59a
0.00b
0.00b
0.34b
0.00b
0.00b
0.17b
5-Methyl-6-hepten-2-one
981
0.46a
0.85a
0.00a
0.44a
0.81a
0.00a
0.28a
1.01a
Myrcene
988
5.07b
0.00c
0.00c
0.00c
0.00c
3.40b
0.20c
0.00c
o-Cymene
1022
0.00d
8.97b
4.61c
10.33a
11.86a
0.00d
11.39a
0.81d
Limonene
1024
9.14c
0.00e
7.98c
0.28e
0.00e
19.81b
4.21d
0.00e
1,8-Cineol
1026
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
γ-terpinene
1054
0.00c
0.00c
0.86a
0.00c
0.00c
0.00c
0.47a
0.00c
cis-sabinene hydrate
1065
0.00c
0.61a
0.00c
0.73a
0.35b
0.00c
0.00c
0.74a
cis-linalool oxide
1067
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
trans-linalool oxide
1084
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Linalool
1095
0.58f
0.99f
0.43f
1.06f
0.63f
0.69f
0.27f
0.48f
Perylene
1102
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
(E)-isocitral
1177
0.63a
0.98a
0.44a
0.00b
0.24b
0.00b
0.77a
0.58a
Myrtenal
1195
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
Neral
1235
31.36c
29.46c
33.28a
27.34c
29.05c
0.00f
27.94c
36.14a
Carvone
1239
5.27d
0.00e
1.06e
0.00e
0.29e
72.73b
0.00e
0.00e
Geraniol
1249
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
Geranial
1264
46.11a
45.12a
49.53a
45.36a
45.21a
0.00f
42.06c
54.63a
Myrtanyl acetate
1324
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
Geranyl acetate
1379
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.64a
β-elemene
1389
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
0.00d
β-caryophyllene
1417
0.39c
0.00c
0.00c
0.00c
0.00c
0.00c
1.20c
0.00c
α-guaiene
1437
0.00d
1.36c
0.00d
2.89a
1.58b
0.00d
0.00d
0.00d
γ-muurolene
1478
0.00c
0.00c
0.26c
0.00c
0.00c
2.57a
0.00c
0.00c
trans-calamenene
1521
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
0.00b
1.24a
Elemol
1548
0.00d
0.00d
1.54b
0.00d
0.00d
0.00d
0.00d
0.00d
Germacrene B
1559
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
0.00c
(E)-nerolidol
1561
0.00e
0.00e
1.54c
0.00e
0.00e
0.00e
0.00e
0.00e
Spathulenol
1577
0.00e
2.85b
0.00e
2.59c
1.82d
0.00e
0.00e
0.00e
Caryophyllene oxide
1582
0.99e
4.10d
0.00e
3.58d
4.59d
0.00e
9.28c
1.57e
Humulene epoxide II
1608
0.00d
1.17a
0.00d
1.33a
0.36c
0.00d
0.71b
0.00d
Essential oil content (%)
1.59c
0.66e
2.80a
1.23d
1.17d
1.95c
1.46d
1.20d
RRI: relative retention index. Means followed by different letters in each row were significantly different by the Scott-Knott test (p<0.05).The variation in the concentrations of compounds among the accessions may have been a consequence of their origin, considering that in this experiment all of the accessions were grown in the same environment. Similar results were observed previously in studies with three chemotypes ofL. alba from different Brazilian states (Rio de Janeiro, Ceará, and São Paulo) [15] and with accessions ofPogostemon sp. from the Active Germplasm Bank of the Federal University of Sergipe [16].The chemical analysis (Table2) indicated that the most abundant compounds among the accessions were 1,8-cineole, linalool, myrcene, limonene, carvone, geranial, and neral, leading to the formation of six clusters according to the obtained chemical compositions, which were differentiated through cluster analysis (Figure 1). The compound linalool, present in accessions LA-01 and LA-22 (84.73% and 84.45%, resp.), showed the highest abundance. A similar result was reported by [17]. Linalool is widely used in the perfume, cosmetic, and fragrance industries [12].Figure 1
Two-dimensional dendrogram showing the similarity of the chemical compositions of 48L. alba accessions obtained from the Active Germplasm Bank at the Federal University of Sergipe.Considering the similarities of the chemical constituents of the essential oils of these 48 accessions, the clusters were classified as follows: Cluster 1 included LA-01, LA-09, LA-22, and LA-27, which contained the following major compounds: linalool, 1,8-cineole, and caryophyllene oxide. Cluster 2 was formed by accessions LA-20 and LA-24 and comprised the following major compounds: linalool, geranial, neral, 1,8-cineole, and caryophyllene oxide. Cluster 3 was formed by accession LA-13, with the following major compounds: limonene, carvone, and sabinene. Cluster 4 comprised accessions LA-56, LA-57, and LA-70, with the following major compounds: carvone, limonene, g-muurolene, and myrcene. Cluster 5 was formed by accessions LA-02, LA-04, LA-08, LA-10, LA-15, LA-17, LA-19, LA-21, LA-28, LA-29, LA-32, LA-37, LA-40, LA-41, LA-42, LA-44, LA-45, and LA-49 and exhibited the following major compounds: neral, geranial, and caryophyllene oxide. Cluster 6 consisted of accessions LA-03, LA-30, LA-36, LA-39, LA-43, LA-52, LA-53, LA-54, LA-55, LA-58, LA-59, LA-60, LA-61, LA-62, LA-63, LA-67, LA-68, LA-69, LA-71, and LA-72, which contained geranial, neral, o-cymene, limonene, and caryophyllene oxide (Figure2).Figure 2
Means and SEM for the chemical compounds in the essential oils from clusters 1 through 6 ofL. alba accessions.
(a)
(b)
(c)
(d)
(e)
(f)According to the PCA (Figure3), the first principal component accounted for 20.55% of the total variability in the data and was positively associated with 1,8-cineol (r=0.82),cis-linalool oxide (r=0.79), linalool (r=0.82), andtrans-linalool oxide (r=0.77) and negatively associated with geranial (r=0.90) and neral (r=-0.89).Figure 3
Distribution of chemical compounds in the essential oils ofL. alba in relation to the first and second principal components, based on the principal component analysis (PCA).The second principal component represented 12.39% of the total variation in the data and showed a positive correlation with 1-octen-3-ol (r=0.60), myrtenal (r=0.69), myrtanyl acetate (r=0.54), and caryophyllene oxide (r=0.75) and a negative correlation with carvone (r=-0.49) and limonene (r=-0.63).


## 4. Conclusions

Variability was observed in the chemical composition of the essential oils ofL. alba accessions obtained from the AGB at UFS, resulting in the formation of six different groups.The compounds characterizing the six groups were as follows: group 1: linalool, 1,8-cineole, and caryophyllene oxide; group 2: linalool, geranial, neral, 1,8-cineol, and caryophyllene oxide; group 3: limonene, carvone, and sabinene; group 4: carvone, limonene, g-muurolene, and myrcene; group 5: neral, geranial, and caryophyllene oxide; and group 6: geranial, neral, o-cymene, limonene, and caryophyllene oxide.The most abundant compounds were 1,8-cineol, linalool, myrcene, limonene, carvone, geranial, and neral.



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*Source: 321924-2015-05-14.xml*