Oil in Tien Giang Province, Vietnam Tan Phat Dao 1,2,3 , Thi Cam Quyen Ngo 1,2 , Thien Hien Tran 1,2 , Thi Kim Ngan Tran 1,2 , Long Giang Bach 1Center of Excellence for Biochemistry and
Trang 1Study on Extraction and Determination of Chemical Composition of Pomelo
(Citrus Grandis L.) Oil in Tien Giang Province, Vietnam
Tan Phat Dao 1,2,3 , Thi Cam Quyen Ngo 1,2 , Thien Hien Tran 1,2 , Thi Kim Ngan Tran 1,2
, Long Giang Bach
1Center of Excellence for Biochemistry and Natural Products, Nguyen Tat Thanh University, Ho
Chi Minh City, Vietnam
*Corresponding author:
Abstract
Hydro-distillation extraction method was used to extract essential oil from pomelo (Citrus Grandis L.) peels in Vietnam, the method is applicable from laboratory scale to pilot scale, production scale.
The effects of extraction variables including size of material, temperature, extraction time, and water-to-material ratio on the yield of essential oil were investigated In addition, to assess the quality of pomelo essential oil, the chemical composition has also been presented in detail The optimized conditions was as follows: material size of grind, temperature of 120oC, ratio of 5 mL/g, and extraction time of 105 min From this parameter, subsequent extraction process related to pomelo oil would a significantly reduce extraction time and improve the quality The chemical composition of the pomelo oil was then determined by GC-MS, and 5 components were identified
In particular, Limonene had the highest content (96.122%) in pomelo essential oil extracted by hydro-distillation extraction This method can be considered as a green method of extraction method as it is less energy intensive process and offers high performance This method is the conventional method but it has great potential for industrial applications and the optimization of pomelo oil extraction conditions is essential
Keywords: Pomelo peels (Citrus Grandis L.), hydro-distillation, GC-MS analysis
1 Introduction
Essential oils are the smallest droplets, concentrated of the herbs, sometimes it is only aromatic molecules formed through the regulatory function in plants It is a mixture of organic substances mixed together, has a characteristic odor, is widely applied in manufacturing and life, especially in the cosmetics, food and pharmaceutical industry [1-13]
Pomelo (Citrus Grandis L.) belongs to genus Citrus, a flowering plant in the Rutaceae family [14].
Pomelo fruit is one of the most popular citrus fruits and is the largest of citrus fruits Currently, pomelo essential oil has been widely applied in many different fields such as cosmetics, perfumes, detergents, aromas and preservatives of food products Pomelo essential oil also has many healing effects and they aid in the distribution of drugs and disinfectants [15] Pomelo essential oil is also used for its bactericidal, antioxidant and anti-cancer properties [16] The chemical composition of Pomelo essential oil from different countries is also a subject of extensive research
Trang 2In this study, hydro-distillation technique was used to extract pomelo essential oils Hydro-distillation does not require a lot of complex equipment, but is capable of almost completely distilling essential oils in raw materials In addition, the method allows the separation of the constituents contained in essential oils into separate parts of higher purity based on differences in volatile properties The volatiles released from the pomelo peel were analyzed by gas
essential oil, while myrcene, sabinene, α-pinene and α-terpinene have been reported at less than 2.5% [17, 18] Other compounds such as: nootkatone, octanal, octyl acetate, citronellyl acetate, citral and carvone have been found at trace levels in essential oil (<1.0%) and have been designated
as the main agents of scents [19], [20] However, we have not found any information on the hydro-distillation process and the composition of essential oil from pomelo peels originating from Tien Giang province, Vietnam Therefore, the study aims to examine the impact of parameters on the extraction process and chemical composition of pomelo oil from Tien Giang province
2 Materials and Methods
2.1 Materials and chemical
Fresh pomelo (Citrus Grandis L.) were selected from Tien Giang province (latitudes 10o25’13’’N
peel were cut into pieces of approximately 2x2 cm2, and kept in an plastic bag at low temperatures (4oC) The sample was weighted exactly and size treated before distillation and was used immediately to avoid the loss of essential oil
2 Hydro-distillation process
Hydro-distillation technique is used to extract pomelo peel essential oil The procedire is as follows: 100g of peels were placed in a 1L round bottomed flask with distilled, deionized water and the essential oil was extracted by hydro-distillation, which was connected to a Clevenger’s distillation unit The pomelo peels was then hydrodistilled for 120 min, the time needed to completely extract essential oils from the tissue [1, 3] The obtained essential oil, which was collected in Clevenger’s, was separated and anhydrous with Na2SO4 The extracted essential oil was place in a glass amber bottle and refrigerated at 4oC until further analysis Each extraction was performed three times
Yield of pomelo peels oil (Y) obtained for every run that was calculated as follow (1):
Y (%) = Volumeofessentialoil (ml)
Amountofrawmaterials (g)100 % (1)
Single factor investigation was used to survey the optimum extraction conditions of the pomelo essential oil and the process was performed four factors: material size, water-material ratio, temperature and extraction time The level of factors is based on single factor experiements with dependent variables as essential oil performance (amount of essential oil in the peels) The survey
is performed to estimate the best conditions for the extraction process, and it was repeated to avoid errors
2.3 Chemical composition analysis by GC-MS
A Gas Chromatography-Mass Spectrometry (GC-MS) is used to analyze the composition of the essential oils of all extraction methods 25 µL sample of essential oil in 1.0 mL n-hexane Name of the equipment: GC Agilent 6890N, MS 5973 inert with HP5-MS column, head column pressure
Trang 39.3psi GC-MS system was performed hold under the following conditions: carrier gas He; flow rate 1.0 mL/min; split 1:100; injection volume 1.0 µL; injection temperature 250oC; oven temperature progress included an initial hold at 50oC for 2 min, and a rise to 80oC at 2oC/min, and them to 150oC at 5oC/min, continue rising to 200oC at 10oC/min and rise to 300oC at 20oC/min for 5 min
3 Results and Discussion
3.1 Single factor investigation
In the extraction process, factors that affect the yield of pomelo essential oil are shown in Figure 1 Figure 1a refers to the change in pomelo oil yield when changing the size of materials during distillation The trend of pomelo essential oil increased as the size of the material decreased, specifically, the efficiency increased from 1% to 1.8% when changing from the original size to pureing the materials This can be explained as follows: the smaller the material size, the faster the water diffuses into the essential oil bag This rapidly pushes the essential oils out under the influence of heat, leading to higher performance On the contrary, the large peels sizes will yield lesser oil in the beginning of the distillation but as have large surfaces to continue to yield more essential oil with increase in distillation time Therefore, grind peels have been chosen in subsequent investigations [21, 22]
One of the main factors influencing the extraction process, was the extraction temperature From Figure 1b, when the temperature increases from 100 to 120oC, the yield of the extracted essential oil gradually increases from 1.5 to 1.8% However, the yield of essential oil nearly no increase at 1.8% and tend to decrease when rising temperature at 130oC and 140oC This observation can be explained from the fact that the heat and the steam generated at 120oC has more rupture effect on the cell structure thus allowing the release of more essential oil at the temperature This result is also found in study of Hien et al [23], the author has announced the efficiency of the process of extracting plants which is greatly affected by temperature Thus for this plant, the temperature of
120oC is the optimal and beyond this temperature, the essential oil will be de degrades and will lose amount due to temperature
Similarly, the effect of extraction time on yield of pomelo essential oil can be seen in Figure 1c At the first stage, extraction time and process yield was directly proportional to each other At the 105 minute time line, yield reaches its highest value at 1.85% and it decreased to 1.75%, 1.6 % at 120 min, 135 min, respectively It can be understood that the longer the extraction time, the more oil is obtained Distillation time extends to a certain limit, the amount of essential oil obtained reaches a state of saturation and is likely to denatured due to prolonged heat exposure [24] The determination of extraction time is related to energy consumption and production cost of the process Therefore, 105 min is used to conduct the next experiment
During distillation, at the boiling point, the essential oils in the plant cells diffuse out the surface of the material and are swept away by the steam The water penetrates the material in the opposite direction and the oil continues to be swept into the water The procedure is carried out until the essential oil in the tissues completely escapes Therefore, determining the ratio of water to raw materials is necessary for the extraction process to reach the maximum amount of essential oil The effect of ratio water and material can be seen in Figure 1d As the ratio changed, the process efficiency also changed and the highest yield was achieved at 5: 1 mL/g At a low water ratio of 2-3 mL/g, the amount of essential oil collected is due to the insufficient amount of water to dissolve the
Trang 4colloidal wrappers surrounding the essential oil bag, the amount of residual essential oil in the material is high, making the efficiency withdrawal of the process On the other hand, at higher ratios such as 6-7:1 mL/g, the amount of essential oil that is obtained is not high, at high rates of water resulting in a slow evaporation rate of the distillation process and possibly essential oils losses due to distillation of water through the collection tube of essential oil, effectively reducing the distillation process In practical terms, large amounts of water lead to prolonged distillation times, costly The highest amount of essential oil obtained (1.9%) at the water-material ratio of 5:1 mL/g
Figure 1 Influence of (A) size of the materials, (B) Temperature, (C) Extraction time, (D)
Water-to-material ratio to the yield of the essential oil
3.2 The results of GC-MS
The compositions of essential oil from pomelo peels was determined by comparing the retention time and mass spectra of the compounds in the essential oil with the spectral data library Citrus essential oils are usually characterized by one or two main compounds
In this study, from 100g of fruit rind around 1.9 mL of essential oil has been obtained by hydro-distillation using Clevenger’s apparatus at optimal conditions The composition of the volatile oil was analyzed by GC-MS (Table 1 and Figure 2) Five constituents were detected from peels, of which, 100% of the compound has been identified Limonene, α-Pinene, β-Myrcene were the main components in the essential oil extracted from pomelo peels was analyzed by GC-MS (Table and
Trang 5figure) Limonene, a monoterpene hydrocarbon, is the most abundant component at 97.1% The other important constituents were β-Myrcene (1.3%) and α-Pinene (0.7%) These results are similar with the other literature data for Citrus genus (Uysal et al., 2011 [14]; Chen et al., 2011 [25]) Essential oils from grapefruit which was reported to be strong antibacterial activity against the bacterial strains (Gram-negative and Gram-positive bacteria), yeast and mold tested by Sumonrat Chanthaphon et al [26] Also, Lis-Halcyon et al [27] identified that Limonene, α-Pinene, and Sabinene exhibit strong anti-bacterial activity by disruption of bacterial or fungal membrane integrity and inhibition of ion transport processes and respiration
Table 1 Chemical constituents of pomelo oil.
Figure 2 The GC-MS chromatogram of essential oil from pomelo peels
4 Conclusion
The extraction of essential oil from pomelo peels using hydro-distillation method is effect by some parameter, such as: water-to-materials ratio, temperature, and extraction time The optimum conditions for extraction process as follows: temperature of 120oC, ratio of 5:1 mL/g, and extracted for 105 minutes Pomelo ssential oil yield of 1.9% in this optimum condition Besides, GC-MS analysis indicates that the composition of pomelo oil in Vietnam obtained using hydro-distillation method contain 97% Limonene Hydro-distillation can be considered a green extraction method because it is a low energy process and does not require a subsequent solvent separation stage Although this method has existed for a long time, it still shows the optimal ability in the extraction
Trang 6process from the experimental scale to the production scale With a compact device, easy to manufacture and simple operation, this method can be applied to extract essential oils from many different material sources and the process of optimizing the extraction process is necessary for each material
Acknowledgments: This study was supported by Nguyen Tat Thanh University, Vietnam
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