Microsoft Word - 10-Agra_48016 1175 Bioscience Journal Original Article Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 INFLUENCE OF STORAGE TEMPERATURES AND STORAGE TIME OF DRY LEAVES ON PATCHOULI [Pogostemon cablin (Blanco) Benth.] ESSENTIAL OIL INFLUÊNCIA DO TEMPO E TEMPERATURAS DE ARMAZENAMENTO DAS FOLHAS SECAS NO ÓLEO ESSENCIAL DE PATCHOULI [Pogostemon cablin (Blanco) Benth.] Maria de Fátima ARRIGONI-BLANK1; Priscila Santana SANTOS1; Arie Fitzgerald BLANK1; Fabiany de Andrade Brito1; Thiago Matos ANDRADE1; Daniela Aparecida de Castro NIZIO1; Péricles Barreto ALVES2 1. Departamento de Engenharia Agronômica, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil, fatima.blank@gmail.com; 2. Departamento de Química, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil ABSTRACT: Patchouli [Pogostemon cablin (Blanco) Benth.] is a plant of the family Lamiaceae, widely used as an essential oil in the cosmetics and perfumery industry. This study aimed to evaluate the influence of storage time and temperature of dry leaves on the patchouli essential oil content and chemical composition. The experiment was performed in a completely randomized design in a 6x2x2 factorial scheme, testing storage time (0, 1, 2, 4, 8, and 16 weeks) and temperature (28°C and 33°C) of dry leaves of two patchouli genotypes (POG-015 and POG-021). The variables essential oil content and chemical composition, and the identification of fungus during storage were evaluated. Results showed that the storage significantly influenced the essential oil content. Patchoulol was identified as the major compound in both genotypes, ranging from 55.05% to 68.77% (POG-15) and from 52.83% to 64.06% (POG-021). Based on the results of patchoulol, dry leaves of both genotypes (POG-015 and POG-021) can be stored for up to eight weeks at 28- 33°C without altering the essential oil quality. KEYWORDS: Pogostemon cablin. Postharvest. Patchoulol. Chemical composition. INTRODUCTION Patchouli [Pogostemon cablin (Blanco) Benth] is an aromatic plant of the family Lamiaceae (ANONIS, 2006) that naturally occurs in Southeast Asia, where it is widely used in traditional medicine (SWAMY; SINNIAH, 2015). The essential oil of this species can be used for several purposes, such as aromatherapy (SALERNO; REBELO; SILVA JUNIOR, 2004), insecticide (PAVELA, 2005), and insect repellent (ZHU et al., 2003; SALERNO; REBELO; SILVA JUNIOR, 2004; ALBUQUERQUE et al., 2013). Patchouli essential oil stands out in the perfumery industry (SINGH; SHARMA; RAMESH, 2002) for providinga base and lasting character to a fragrance, and is among the 18 most important essential oils worldwide (BIZZO; HOVELL; REZENDE, 2009). The chemical composition of patchouli essential oil varies according to the genotype, production organs, environmental and production conditions, and storage conditions. However, patchoulol and α-patchoulene are considered the most important compounds and aroma regulators (SINGH; SHARMA; RAMESH, 2002). Patchoulol is a sesquiterpene officially used as a chemical marker for the quality assessment of Pogostemonis Herba in the Chinese Pharmacopoeia. Tsai et al. (2007) reported that leaf material quality, dry leaf, essential oil aging, and plant age could also influence the quality of patchouli essential oil. A review study has shown that when considering 100 analyses of patchouli essential oils, the authors listed the main compounds and their observed average concentrations, such as patchoulol (39.0%), α-bulnesene (14.0%), α-guaiene (11.0%), seyclellene (6.6%), α-patchoulene (4.5%), (E) -β- caryophyllene (3.1%), β-patchoulene and pogostol (2.4%) (BEEK; JOULAIN 2018). Nizio et al. (2018) reported that, in general, the stages that follow the plant material harvest, such as the drying process, the storage of plant material, and the extraction process could influence the essential oil content and chemical composition. Leaf drying and the appropriate storage allow better plant material conservation. Thus, the drying process is widely used, but it must be performed in a way that does not compromise the product quality at the end of the production process (MARCHESE; Received: 08/04/19 Accepted: 20/12/19 1176 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 FIGUEIRA, 2005). In a study with Myrcia lundiana, Alves et al. (2018) reported that one-day drying at 40 ° C was enough to dry the leaves without affecting the essential oil content. In the case of aromatic species, the drying process must consider the storage structure and the volatile characteristics of the essential oil compounds. Drying is used to prevent microorganisms reproduction, consequently preserving the quality of the harvested plant. It also reduces the weight and volume of plants for transportation and storage (CORREA et al., 2004). Sant'ana et al. (2010) observed that the dry leaves of two patchouli genotypes stored for 28 days had an increase in the essential oil content. The novelty of the present study is the storage time and the comparison of the percentages of the main compounds at those storage times. Due to the importance of patchouli essential oil, this study aimed to evaluate the effect of the storage time and temperature of dry leaves on the essential oil content and chemical composition. MATERIAL AND METHODS Plant material and experimental design This study used two patchouli genotypes (POG-015 and POG-021) from the Active Germplasm Bank of Medicinal and Aromatic Plants of the Federal University of Sergipe. The exsiccates are deposited in the Herbarium of the Department of Biology, under the numbers 13173 and 13177. Seedlings of the two genotypes were generated by cutting, in black plastic bags. Vermiculite and coconut coir dust (1: 1) were used as the substrate and were supplemented with 1g.L-1 of lime and 2g.L-1 of N-P-K (6 -24-12). Seedlings were transferred to a greenhouse (50% shaded), with an intermittent misting, at the Department of Agronomic Engineering of UFS, where they were kept until transplanting. At 40 days after planting, seedlings were transplanted to the field (January 2011). Seedlings were planted at the Experimental Farm "Campus Rural da UFS", located in the municipality of São Cristóvão, Sergipe (lat. 11°00'S; long. 37°12'W), in a Red-Yeallow Argisol. The soil had the following chemical and physical characteristics: water pH - 5.4; P - 23.3mg.dm-3; K - 0.09 cmolc.dm-3; Ca + Mg - 2.70 cmolc.dm-3; Al = 0.48 cmolc.dm-3; Zn - 1.09 cmolc.dm-3; Cu - 0.68 cmolc.dm-3; Fe - 990.0 cmolc.dm-3; Mn - 1.14 mg.dm-3; O.M - 21 g.dm-3; and V - 58.1%. The soil pH was corrected to 6.5, using dolomitic lime. Fertilization consisted of applying 6L.m-2 of bovine manure and 160g.m-2 of N-P-K (6- 24-12) + micronutrients. The spacing was 0.50m between plants and 0.60m between rows. Drip irrigation was used, and plants were cultivated in double-colored plastic mulch (black on the bottom and white on the top). In May 2011, shoots were harvested at 25 cm from the soil, and leaves were dried in a forced- air-circulation oven at 38 °C, for five days. Then, the shoot dry matter per plant was determined. Dry leaves were randomly separated into 60-gram lots and packed in black plastic bags. Leaves were stored in a BOD chamber, in the darkness, for 0, 1, 2, 4, 8, and 16 weeks, at 28 and 33 °C. The experiment was carried out in a completely randomized design, in a 6x2x2 factorial scheme, with six storage periods (0, 1, 2, 4, 8, and 16 weeks), two Genotypes (POG-015 and POG- 021), and two temperatures (28 and 33 °C), with three replications. Essential oil extraction Three 50g dry leaves samples were collected in the storage period for the essential oil extraction. The metabolite was extracted by hydrodistillation, in a Clevenger apparatus coupled to a 3,000mL round-bottomed glass flask, for 160 minutes. The flask was added with 2,000mL of distilled water and 50g of dry leaves (EHLERT et al., 2006). Contents were estimated based on the dry mass (mL.100g-1) and expressed in percentage. Essential oil samples were kept in capped amber glass flasks, in a freezer, until chemical analysis. Essential oil chemical analysis The qualitative analysis of the essential oil chemical composition was performed in a gas chromatograph coupled to a GC-MS mass spectrometer (Shimadzu, model QP 5050A) with an autosampler (AOC-20i, Shimadzu), and a J & W Scientific fused silica capillary column (5%- diphenyl-95%-dimethylpolysiloxane) of 30 m x 0.25 mm i.d., 0.25 μm film thickness, at a constant helium flow rate of 1.2 mL/min. The temperature was set to 50◦C for 1.5 min, then programmed to increase to 200◦C at 4◦C/min, followed by an increase of 15◦C/min up to 250◦C, then remaining constant for 5 min. The sampler temperature was 250° C, and the detector temperature was 280 °C. The injection volume was 0.5μL (ethyl acetate), with a split ratio of 1: 100 and a column pressure of 64.20kPa. The MS conditions consisted of an ionic capture detector with an electron energy of 70eV. The scanning speed was 1,000, at a scan rate of 0.3 scan/s, and the system was programmed to scan Received: 08/04/19 Accepted: 20/12/19 1177 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 fragments/ions with m/z in the order of 40 to 500 Da. The essential oil components were identified by matching their mass spectra to the spectra available in the database of the device (NIST05, NIST21, and WILEY8). These libraries allowed for the comparison of spectral data with a similarity index above 80%. The measured retention indices were also compared with indices from the literature (ADAMS, 2007). The Kovats retention indices (KI) were determined by a homologous series of n-alkanes (C8-C18) injected under the same chromatographic conditions of the samples, using the equation of Van Den Dool and Kratz (1963). The quantitative analysis of the essential oil compounds was performed by Gas Chromatography/Mass Spectrometry and Flame Ionization Detector - GC/MS/FID (Shimadzu GC- 17A), coupled with a ZB-5MS fused silica capillary column (5% dimethylpolysiloxane), with 30m x 0.25mm i.d., 0.25μm film thickness, under the same GC-MS conditions. Compounds were quantified by area normalization (%). Compounds concentrations were calculated by the area and placed in order of GC elution. Statistical analyses Variables means were subject to analysis of variance and compared by the Scott-Knott test at the 5% probability level and polynomial regression, using the Sisvar software (FERREIRA, 2011). RESULTS AND DISCUSSION A triple interaction (genotype x storage time x storage temperature of leaves) was detected in the essential oil content. The essential oil content of genotype POG-021 varied when the leaves were stored at 28 °C. This variation is represented by a quadratic equation, in which the highest levels were obtained between the second and eighth weeks of storage. At this same temperature (28°C), no significant differences were observed for genotype POG-015. At 33°C results for both genotypes can be represented by quadratic equations, and contents ranged from 2.13% to 3.0% (POG-015) and from 2.43% to 3.0 % (POG-021) (Table 1). Table 1. Essential oil content of Patchouli (Pogostemon cablin) genotypes in function of storage time and storage temperature of dry leaves. Genotype Essential oil content (%) Time (weeks) 0 1 2 4 8 16 Equation Y = ------------------------------------- 28°C --------------------------------- POG-015 2.40aA 3.27aA 2.60bA 2.60bA 2.73bA 2.30bA ns POG-021 2.60aA 3.07aA 3.60aA 3.47aA 3.60aA 2.93aA 2.8431+0.2197x-0.0135x 2 R2= 73.95% ------------------------------------- 33°C---------------------------------- POG-015 2.40aA 2.87aA 2.87aA 2.73aA 3.00aA 2.13aA 2.556+0.1262x-0.0095x2 R2=81.16 POG-021 2.60aA 3.00aA 2.87aB 2.93aB 3.00aB 2.43aA 2.7303+0.0879x-0.0066x2 R2= 80.30% CV (%) 11.21 São Cristóvão (SE), UFS, 2012; Means followed by the same lowercase letters in the column and uppercase letter between temperatures do not differ by the Scott-Knott's test (p≤0.05). Sant'ana et al. (2010) reported the beneficial effect on the essential oil content provided by the storage of patchouli leaves for 28 days. The authors verified an increase from 1.60 to 2.7% in the essential oil content, depending on the genotype. They also detected 14 chemical compounds in two patchouli genotypes. The highest percentages of essential oil compounds in the two storage temperatures were reported for patchoulol, with values ranging from 52.04 to 68.77% (POG-015) and from 52.83 to 64.06% (POG-021); α-bulnesene, with values ranging from 6.48 to 10.20% (POG-015) and from 5.71 to 9.67% (POG-021); pogostol, with values ranging from 5.45 to 7.72% (POG-015); and α- guanene, with values ranging from 4.14 to 7.20% (POG-015) and from 5.30 to 7.16% (POG-021). Considering β-patchoulene, the highest content (1.46%) reported for genotype POG-015 was obtained around the fourth week of leaves storage at 28°C. Conversely, for genotype POG- 021, at the same temperature, the highest content (1.85%) was recorded in the sixteenth week of storage. The analysis of genotype POG-021 at 33°C 1178 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 showed no significant differences throughout the storage period. The same was observed for β- elemene and cycloseychellene (Table 2). E-caryophyllene contents ranged from 1.07% in the eighth week to 2.10% in the second week (POG-015, at 28°C). For POG-021, the contents varied between 1.30% (eighth week) and 2.06% (first week) at 33°C. At 28°C, results for both genotypes are represented by cubic equations (Table 3). In relation to α-guanene, contents were slightly higher when compared with those of E-caryophyllene, ranging from 4.14 to 7.20% (POG-015). For seychellene, the lowest contents were recorded in the eighth week for POG-015, at the two temperatures tested; conversely, the highest contents were registered for POG-021, in the sixteenth week, with no significant difference between the two temperatures (Table 3). Table 2. Contents of the chemical compounds β-patchoulene, β-elemene, and cycloseychellene of the essential oil of Patchouli (Pogostemon cablin) genotypes in function of storage time and storage temperatures of dry leaves. Genotype Storage (weeks) 0 1 2 4 8 16 Equation Y = β-patchoulene (RRI: 1379) ------------------------------------- 28°C--------------------------------- POG-015 1.13aA 1.14aA 1.17Aa 1.46aA 0.77aA 1.12bA 1.0354+0.2282x- 0.0501x2+0.0022x3 R2= 73.38% POG-021 1.34aA 1.08aB 1.24aA 0.98bA 1.08aA 1.85aA 1.2892-0.0949x+0.0081x2 R2= 93.38% ------------------------------------- 33°C--------------------------------- POG-015 1.13aA 1.29bA 1.20aA 1.07aB 0.80bA 1.08bA 1.2752-0.0820x+0.0042x2 R2=65.97% POG-021 1.34aA 1.76aA 1.36aA 1.07aA 1.40aA 1.46aB Ns CV (%) 16.23 β-elemene (RRI: 1389) ------------------------------------- 28°C--------------------------------- POG-015 0.57aA 0.60aA 0.63aA 0.62aA 0.30aA 0.38aA 0.5525+0.0899x- 0.0238x2+0.0010x3 R2= 98.97% POG-021 0.43bA 0.46bA 0.37bA 0.49bA 0.32aA 0.44aA ns ------------------------------------- 33°C--------------------------------- POG-015 0.57aA 0.58aA 0.64aA 0.49aB 0.33aA 0.38aA 0.6286-0.0460x+0.0018x2 R2= 79.51% POG-021 0.43bA 0.51bA 0.36bA 0.44aA 0.31aA 0.39aA ns CV (%) 9.01 Cycloseychellene (RRI: 1406) ------------------------------------- 28°C--------------------------------- POG-015 0.26aA 0.28aA 0.29aA 0.34aA 0.46aA 0.23bA 0.2333+0.0471x-0.0029x2 R2= 86.87% POG-021 0.26aA 0.31aA 0.33aA 0.31aA 0.34bA 0.34aA ns --------------------------------- 33°C --------------------------------- POG-015 0.26aA 0.27aA 0.30aA 0.26aA 0.26aB 0.22aA ns POG-021 0.26aA 0.34aA 0.25aA 0.33aA 0.25aA 0.30aA ns CV (%) 18.05 São Cristóvão (SE), UFS, 2012; Means followed by the same lowercase letters in each column and uppercase letters between temperatures do not differ by the Scott-Knott's test (p≤0.05). The contents of α-humulene, allo- aromadendrene (Table 4), and 9-epi- (E) caryophyllene (Table 6) were below 1.0%. The content of α-patchoulene decreased from the eighth week of storage for POG-015 at both temperatures. POG-021 had no significant difference for storage time at 28 and 33 °C (Table 4). α-bulnesene contents ranged from 6.5 to 10% (POG-015) and from 5.7 to 9.6% (POG-021) (Table 5). The contents of α-bulnesene in POG-015 increased until the fourth week of storage at 28°C. The highest patchoulol content in POG-015 was reported in the eighth week of storage (68.77%) at 28 °C. Results are represented by cubic equations 1179 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 for the two genotypes (Table 6). Sant'ana et al. (2010) also observed that patchoulol concentrations of the essential oil of stored leaves were significantly higher than those of non-stored leaves. Table 3. Contents of the chemical compounds E-caryophyllene, α-guaiene, and seychellene of the essential oil of Patchouli (Pogostemon cablin) genotypes in function of storage time and storage temperatures of dry leaves. Genotype Storage (weeks) 0 1 2 4 8 16 Equation Y = E-caryophyllene (RRI: 1417) ------------------------------------- 28°C--------------------------------- POG-015 1.89aA 2.02aA 2.10aA 2.13aA 1.07bA 1.23bA 1.8346+0.3115x- 0.0791x2+0.0035x3 R2= 98.37% POG-021 1.67aA 1.85aA 1.74bA 1.86bA 1.48aA 1.73aA 1.6801+0.1341x- 0.0312x2+0.0014x3 R2= 82.56% ------------------------------------- 33°C--------------------------------- POG-015 1.89aA 1.92aA 2.04aA 1.71aB 1.09aA 1.14bA 2.0713-0.1391x+0.0049x2 R2= 83.98% POG-021 1.67aA 2.06aA 1.58aB 1.85aA 1.30aA 1.47aB 1.7357+0.1142x- 0.0335x2+0.0015x3 R2= 62.05% CV (%) 8.88 α-guaiene (RRI: 1437) ------------------------------------- 28°C--------------------------------- POG-015 6.38aA 6.97aA 6.80aA 7.20aA 4.20bA 4.62bA 6.2763+0.8599x- 0.2170x2+0.0098x3 R2= 96.50% POG-021 5.87aA 6.78aA 6.83aA 6.96aA 6.14aA 7.03aA 5.9908+0.6865x- 0.1290x2+0.0056x3 R2= 93.65% ------------------------------------- 33°C--------------------------------- POG-015 6.38aA 6.47aA 6.75aA 6.02aA 4.14bA 4.59bA 21 5.87aA 7.16aA 6.21aA 6.90aA 5.30aA 6.18aA 6.8955- 0.3933x+0.0151x2 R2= 80.62% 6.0567+0.6252x- 0.1405x2+0.0063x3 R2= 65.53% CV (%) 9.53 Seychellene (RRI: 1444) ------------------------------------- 28°C--------------------------------- POG-015 3.70aA 4.59aA 3.91aA 4.14aA 2.88bA 3.32bA 3.8610+0.3803x- 0.1000x2+0.0046x3 R2= 81.29% POG-021 3.69aA 3.88bA 4.14aA 3.97aA 4.08aA 4.72aA Ns ------------------------------------- 33°C--------------------------------- POG-015 3.70aA 3.78bB 3.95aA 3.48bB 2.95bA 3.33bA 3.9316-0.1595x+0.0074x2 R2= 71.13% POG-021 3.69aA 4.35aA 3.83aA 4.03aA 3.75aA 4.21aA Ns CV (%) 8.53 São Cristóvão (SE), UFS, 2012; Means followed by the same lowercase letters in the column and uppercase letters between temperatures do not differ by the Scott-Knott's test (p≤0.05). 1180 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 Table 4. Contents of the chemical compounds α-humulene, α-patchoulene, and allo-aromadendrene of the essential oil of Patchouli (Pogostemon cablin) genotypes in function of storage time and storage temperatures of dry leaves. Genotype Storage (weeks) 0 1 2 4 8 16 Equation Y = α-humulene (RRI: 1452) ------------------------------------- 28°C--------------------------------- POG-015 0.41aA 0.45aA 0.46aA 0.46aA 0.28aA 0.27bA 0.4060+0.0573x- 0.0141x2+0.0006x3 R2= 99.71% POG-021 0.34bA 0.38bA 0.38bA 0.38bA 0.34aA 0.36aA 0.3470+0.0274x- 0.0055x2+0.0002x3 R2= 86.38% ------------------------------------- 33°C--------------------------------- POG-015 0.41aA 0.42aA 0.46aA 0.37aB 0.26bA 0.26aA 0.4487-0.0270x+0.0009x2 R2= 82.07% POG-021 0.34bA 0.43aA 0.34bA 0.37aA 0.33aA 0.31aA Ns CV (%) 9,52 α-patchoulene (RRI: 1454) ------------------------------------- 28°C--------------------------------- POG-015 2.44aA 2.61aA 2.59aA 2.82aA 1.78bA 1.97bA 2.3789+0.3246x- 0.0769x2+0.0034x3 R2= 93.49% POG-021 2.41aA 2.59aB 2.75aA 2.53aA 2.62aA 2.89aA Ns ------------------------------------- 33°C--------------------------------- POG-015 2.44aA 2.51bA 2.66aA 2.34bB 1.79bA 1.98bA 2.6358-0.1216x+0.0048x2 R2= 73.76% POG-021 2.41aA 2.94aA 2.50aA 2.69aA 2.24aB 2.52aB Ns CV (%) 8.54 Allo-aromadendrene (RRI: 1458) ------------------------------------- 28°C--------------------------------- POG-015 0.59bA 0.48bA 0.48bA 0.48bA 0.56bA 0.61bA 0.5693 0.0707x+0.0128x2+0.0005x3 R2= 90.18% POG-021 0.83aA 0.91aA 0.90aA 0.78aA 0.78aA 0.97aA 0.8936-0.0298x+0.0021x2 R2= 68.07% ------------------------------------- 33°C--------------------------------- POG-015 0.59bA 0.55bA 0.55bA 0.55bA 0.55bA 0.61bA Ns POG-021 0.83aA 1.02aA 0.82aA 0.87aA 0.76aA 0.81aB Ns CV (%) 11.38 São Cristóvão (SE), UFS, 2012; Means followed by the same lowercase letters in each column and uppercase letters between temperatures do not differ by the Scott-Knott's test (p≤0.05). The highest content of pogostol (7.72%) in POG-015 was obtained in the second week of storage at 33 °C. Results are represented by a cubic equation (Table 6). The compounds found in this study agree with what is commonly observed for patchouli essential oil. According to a review published exclusively with this aromatic species, the three compounds observed at the highest concentrations for most published studies are patchoulol, α- bulnesene, and α-guaiene (BEEK; JOULAIN 2018). The storage time and the storage temperature influence the patchouli essential oil content. The highest essential oil content of genotype POG-021 leaves was detected between the second and eighth weeks of storage at 28 °C. Based on patchoulol, dry leaves of patchouli genotypes (POG-015 and POG-021) can be stored for up to eight weeks at 28 °C and 33 °C without altering the essential oil quality 1181 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 Table 5. Contents of the chemical compounds 9-epi- (E) caryophyllene and α-bulnesene of the essential oil of Patchouli (Pogostemon cablin) genotypes in function of storage time and storage temperatures of dry leaves. Genotype Storage (weeks) 0 1 2 4 8 16 Equation Y = 9-epi-(E) caryophyllene (RRI: 1464) ------------------------------------- 28°C------------------------------------- POG-015 0.78aA 0.86aA 0.85aA 0.89aA 0.55bA 0.60bA 0.7677+0.1065x- 0.0257x2+0.0011x3 R2= 97.24% POG-021 0.69aA 0.76aA 0.78aA 0.76bA 0.74aA 0.82aA Ns ------------------------------------- 33°C------------------------------------- POG-015 0.78aA 0.79aA 0.85aA 0.74aB 0.52bA 0.59bA 0.8470- 0.0466x+0.0018x2 R2= 74.54% POG-021 0.69aA 0.85aA 0.70bA 0.79aA 0.64aA 0.73aA Ns CV (%) 8.73 α-bulnesene (RRI: 1509) ------------------------------------- 28°C------------------------------------- POG-015 9.27aA 10.00aA 10.20aA 10.17aA 6.78aA 7.16aA 9.1980+1.0736x- 0.2674x2+0.0120x3 R2= 99.30% POG-021 5.71bA 8.55aA 9.18aA 9.51aA 8.54aA 9.67aA 6.1477+2.0627x- 0.3302x2++0.0134x3 R2= 91.85% ------------------------------------- 33°C------------------------------------- POG-015 9.27aA 9.49aA 9.84aA 6.48aB 7.09aA 7.30aA 9.7909- 0.6210x+0.0292x2 R2= 66.06% POG-021 5.71bA 9.37aA 8.70aA 9.06aA 7.80aA 8.58aA 6.4646+1.8311x- 0.3157x2+0.0131x3 R2= 68.05% CV (%) 19.83 São Cristóvão (SE), UFS, 2012; Means followed by the same lowercase letters in the column and uppercase letters between temperatures do not differ by the Scott-Knott's test (p≤0.05). Table 6. Contents of the chemical compounds caryophyllene oxide, pogostol, and patchoulol of the essential oil of Patchouli (Pogostemon cablin) genotypes in function of storage time and storage temperatures of dry leaves. Genotype Storage (weeks) 0 1 2 4 8 16 Equation Y = Caryophyllene oxide (RRI: 1582) ------------------------------------- 28°C------------------------------------- POG-015 0.65aA 0.61aA 0.73aB 0.79aB 0.70bA 1.37aB 0.6817- 0.0139x+0.0034x2 R2= 92.52% POG-021 0.38bA 0.83aA 0.65aA 0.87aB 1.01aA 1.58aB 0.5593+0.0632x R2= 89.64% ------------------------------------- 33°C------------------------------------- POG-015 0.65aA 0.72bA 1.26aA 1.23aA 0.72bA 1.71bA 0.5701+0.4265- 0.0793x2+0.0035x3 R2= 92.62% POG-021 0.38bA 1.01aA 0.63bA 1.36aA 0.99aA 2.06aA 0.4213+0.3949x- 0.0614x2+0.0026x3 R2= 87.24% CV (%) 15.94 Pogostol (RRI: 1651) ------------------------------------- 28°C------------------------------------- 1182 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 POG-015 7.12aA 6.94aA 7.52aA 6.43aB 5.45aA 5.79aA 7.4294- 0.3169x+0.0131x2 R2= 79.32% POG-021 4.44bA 5.45bA 4.36bA 4.58bB 4.47bA 4.98bA Ns ------------------------------------- 33°C------------------------------------- POG-015 7.12aA 7.42aA 7.72aA 7.39aA 5.72aA 6.14aA 7.0946+0.5327x- 0.1391x2+0.0063x3 R2= 99.63% POG-021 4.44bA 5.76bA 4.68bA 5.62bA 4.99bA 5.18bA Ns CV (%) 6.60 Patchoulol (RRI: 1656) ------------------------------------- 28°C------------------------------------- POG-015 55.05bA 53.95aA 52.59bA 52.33bA 68.77aA 62.87aA 56.0921- 4.4147x+1.1841x2- 0.0551x3 R2= 97.36% POG-021 63.49aA 57.34aA 60.26aA 58.91aA 62.53bA 54.24bA 62.2566- 2.6939x+0.5504x2- 0.0258x3 R2= 80.97% ------------------------------------- 33°C------------------------------------- POG-015 55.05bA 54.31aA 52.04bA 56.01aA 67.99aA 61.38aA 55.3201- 2.7955x+0.8975x2- 0.0437x3 R2= 98.90% POG-021 63.49aA 52.83aB 62.31aA 55.41aA 64.06aA 56.92bA Ns CV (%) 4.27 São Cristóvão (SE), UFS, 2012; Means followed by the same lowercase letters in each column and uppercase letters between temperatures do not differ by the Scott-Knott's test (p≤0.05). RESUMO: Patchouli [Pogostemon cablin (Blanco) Benth.] é uma planta pertencente à família Lamiaceae, e seu óleo essencial é utilizado nas indústrias de perfumes e cosméticos. O objetivo do trabalho foi avaliar a influência do tempo e temperaturas de armazenamento das folhas secas no teor e na composição química do óleo essencial de patchouli. O delineamento experimental foi o inteiramente casualizado em esquema fatorial 6x2x2, testando tempo de armazenamento (0, 1, 2, 4, 8 e 16 semanas), temperatura de armazenamento (28°C e 33°C) de folhas secas de dois genótipos (POG-015 e POG-021) de patchouli. As variáveis avaliadas foram o teor e a composição química do óleo essencial e a identificação dos fungos que se desenvolveram durante o armazenamento. Os resultados mostraram que o armazenamento influenciou significativamente o teor de óleo essencial. Dos compostos identificados, o patchoulol foi o composto majoritário nos dois genótipos, variando de 55,05% a 68,77% (POG-15) e 52,83% a 64,06% (POG-021). Baseado no patchoulol, folhas secas dos genótipos (POG-015 e POG-021) de patchouli podem ser armazenadas por um período de até oito semanas em temperatura de 28 e 33°C, sem alterar a qualidade do óleo essencial. PALAVRAS-CHAVE: Pogostemon cablin. Pós-colheita. Patchoulol. Composição química, REFERENCES ADAMS, R. P. Identification of essential oil components by gas chromatography/mass spectroscopy. 4th ed., Allured: Carol Stream, 2007. 804p. ALBUQUERQUE, E. L. D.; LIMA, J. K. A.; SOUZA, F. H. O.; INDIRA M.A. SILVA, I. M. A.; SANTOS, A. A.; ARAÚJO, A. P. A.; BLANK, A. F.; LIMA, R. N.; ALVES, P. B.; BACCI, L. Insecticidal and repellence activity of the essential oil of Pogostemon cablin against urban ants species. Acta Tropica, v. 127 p. 181– 186, apr. 2013. http://dx.doi.org/10.1016/j.actatropica.2013.04.011 1183 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 ALVES, M. F.; BLANK, A. F.; ANDRADE, T. M.; NIZIO, D. A. C.; SAMPAIO, T. S.; NASCIMENTO JUNIOR, A. F. content and chemical composition of the essential oil of Myrcia lundiana kiaersk in different drying times. Biosciene Journal, Uberlândia, v. 34, n. 3, p. 623-628, may/june 2018. http://dx.doi.org/10.14393/BJ-v34n3a2018-39371 ANONIS, D. P. Woody notes in perfumery, patchouly oil, absolute and aroma chemicals: Part I. Perfumer & Flavorist, v. 31, p. 36-39, nov. 2006. Disponível< http://www.media.allured.com/documents/PF_31_11_036_04.pdf> Acesso em: 12 de agosto de 2013. BEEK, V. T. A.; JOULAIN D. The essential oil of patchouli, Pogostemon cablin: A review. Flavour and Fragrance Journal, v. 33, p. 6–51, 2018. https://doi.org/10.1002/ffj.3418 BIZZO, H. R.; HOVELL, A. M. C.; REZENDE, C. M. Óleos essenciais no Brasil: aspectos gerais, desenvolvimento e perspectivas. Química Nova, São Paulo, v. 32, p. 588-594, apr. 2009. http://dx.doi.org/10.1590/S0100-40422009000300005 CORREA, R. M.; BERTOLUCCI, S. K. V.; PINTO, J. E. B. P.; REIS, E. S.; ALVES, T. L. Teor de óleo essencial e caracterização organoléptica de folhas de assa-peixe submetidas a diferentes métodos de secagem. Ciência e Agrotecnologia, Lavras, v. 28, n. 2, p. 339-344, mar/apr.2004. http://dx.doi.org/10.1590/S1413- 70542004000200013. EHLERT, P. A. D.; BLANK, A. F.; ARRIGONI-BLANK, M. F.; PAULA, J. W. A.; CAMPOS, D. A.; ALVIANO, C. S. Tempo de hidrodestilação na extração de óleo essencial de sete espécies de plantas medicinais. Revista Brasileira de Plantas Medicinais, Botucatu, v. 8, n. 2, p. 79-80, 2006. http://ri.ufs.br/handle/123456789/1548 FERREIRA, D. F. Sisvar: a computer statistical analysis system. Ciência e Agrotecnologia, Lavras, v. 35, n. 6, p.1039-1042, nov/dec. 2011. http://dx.doi.org/10.1590/S1413-70542011000600001 MARCHESE, J. A., FIGUEIRA, G. M. O uso de tecnologias pré e pós-colheita e boas práticas agrícolas na produção de plantas medicinais e aromáticas. Revista Brasileira de Plantas Medicinais, Botucatu, v. 7, n. 3, p. 86-96, 2005. http://repositorio.unicamp.br/jspui/bitstream/REPOSIP/93640/1/2-s2.0-23844495644.pdf NIZIO, D. A. C.; BLANK, A. F.; SAMPAIO, T. S.; BRITO, F. A.; ANDRADE, T. M.; ARRIGONI-BLANK, M. F.; MARIA, A. N. Distillation methods affect the chemical composition of Varronia curassavica Jacq. essential oil? Bioscience Journal, Uberlândia, v. 34, n. 3, p. 629-639, may/june 2018. http://dx.doi.org/10.14393/BJ-v34n3a2018-39372 PAVELA, R. Insecticidal activity of some essential oils against larvae of Spodoptera littoralis. Fitoterapia, v. 76, p. 691-696, oct. 2005. http://dx.doi.org/10.1016/j.fitote.2005.06.001 SALERNO, A. R.; REBELO, A. M.; SILVA JUNIOR, A. A. Plantas aromáticas para cultivo em Santa Catarina. Agropecuária Catarinense, Florianópolis, v. 17, n. 2, p. 46-49, 2004. SANT’ANA, T. P. C.; BLANK, A. F; VIEIRA, S. D; ARRIGONI-BLANK, M. F. Influência do armazenamento de folhas secas no óleo essencial de patchouli (Pogostemon cablin Benth.). Química Nova, São Paulo, v. 33, n. 6, p. 1263-1265, jun. 2010. http://dx.doi.org/10.1590/S0100-40422010000600008. SINGH, M.; SHARMA, S.; RAMESH, S. Herbage, oil yield and oil quality of patchouli [Pogostemon cablin (Blanco) Benth.] influenced by irrigation, organic mulch and nitrogen application in semi-arid tropical climate. Industrial Crops and Products, v. 16, n. 2, p. 101-107, feb. 2002. https://doi.org/10.1016/S0926- 6690(02)00013-4 1184 Influence of storage… ARRIGONI-BLANK, M. F. et al. Biosci. J., Uberlândia, v. 36, n. 4, p. 1175-1184, July/Aug. 2020 http://dx.doi.org/10.14393/BJ-v36n4a2020-48016 SWAMY, M. K.; SINNIAH, A. R. “A comprehensive review on the phytochemical constituents and pharmacological activities of Pogostemon cablin Benth. An aromatic medicinal plant of industrial importance”. Molecules, v. 20, n. 5, p. 8521–8547, may 2015. http://dx.doi.org/10.3390/molecules20058521. TSAI, Y.; HSU, H.; YANG, W.; TSAI, W.; CHEN, C.; WATANABE, T. α-bulnesene, a PAF inhibitor isolated from the essential oil of Pogostemon cablin. Fitoterapia, v. 78, p. 7-11, jan. 2007. https://doi.org/10.1016/j.fitote.2006.09.016 VAN DEN DOOL, H.; KRATZ, P. D. A generalization of the retention index system including linear temperature programmed gas-liquid partition chromatography. Journal of Chromatography, USA, v. 11, p. 463-471, aug. 1963. https://doi.org/10.1016/S0021-9673(01)80947-X ZHU, B. C. R.; HENDERSON, G.; YU, Y.; LAINE, R. A. Toxicity and repellency of patchouli oil and patchouli alcohol against formos an subterranean termites Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Journal Agricultural and Food Chemistry, v. 51, p. 4585-4588, july 2003. https://doi.org/10.1021/jf0301495