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will increase the oil recovery, improve oil quality and stability. The essential oil was extracted using the enzyme pectinase. A control and single treatment were manipulated. The freshly harvested lemongrass were processed, then the enzyme (pectinase = 0.5% of the sample+ buffer ) was added to the treatment while only the buffer was added to control (1:6 sample: buffer dilution ratio). After 12 hours of incubation at room temperature with constant shaking, the reaction mixtures were boiled in a water bath to inactivate the enzyme. When cooled, the mixtures were filtered in cheesecloth and extracted with a solvent. The collected organic layer was subjected to the rotary evaporator or electromantel oil extraction evaporator to recover the essential oil. The oil was then filtered busing filter paper. An essential oil yield of 0.96% f the raw sample was obtained for the treatment and 0.47% oil yield for the control. Prior to the physico-chemical analysis, the enzyme-treated lemongrass oil can be considered similar to the of an extra grade oil having a specific gravity of 1.033), solubility in 70% ethanol of 10:1 ratio and a strong lemon-like odor as perceived by the panelists. The untreated sample was of low quality oil (based on the related literature). Given the results, it may be concluded that essential oil production through pectinase application could be answer to the country’s high dependenc on imported essence.
CHAPTER 1 THE PROBLEM AND ITS BACKGROUND Introduction Essential oils are very much in demand in the Philippines, especially in the pharmaceutical and cosmetic industries. These industries heavily depend on foreign essences and cosmetic preparation for equate supply and are spending millions of dollars for the import of essential oil and other toilet Preparation (Manalo et al.,1982). The Philippines rank 8th among the importing countries, garnering 1.3% of the world’s share and practically Imports more than 90% of the countries requirements (Dar, 1997).
the use of pectic enzyme in various industries is increasing. 2.3 million in 1993 while exports Decreased from a value of US$ 2. have a low cost effectivity. it aims: 1. and 4. This study therefore was conducted in order to develop a new technique. . one will assume that the demand for essential oils is also likely to grow. It is also use for the isolation of citral which is employed in artificial flavors and in the manufacture of ionones. 1997). Moreover.5 in 1993.2 million in 1989 to US$ 59. in extracting essential oils from lemongrass so as to increase its yield. in terms of quality (net kg) from 328. To extract essential oils from lemongrass through enzymatic process under optimized condition.4 million in 1989 to US$ 1. 3. It is therefore timely and relevant to further develop and explore the essential oil bearing plants to lessen the county’s dependence on imported essential oils (Jamilla. 1997). as more money become available for discretionary funding.084 kg in 1993 (Jamilla. Given the outlook for the continuing economic growth of the country. expression by hand or machine and enfleurage.High dependence of the Philippines on imported essential oil increased From US $ 37. However. Data for the period 1989-1993 show an increasing trend value of importation of essential oils (De Guzman. To compare the essential oils yield between the control and enzymetreated samples. Specifically. On the other hand. The Research Objectives This study focuses on the production of essential oils from lemongrass through enzyme process. bath salts. To conduct physico-chemical analysis of the extracted essential oils.494 kg in 1989 to 180. Lemongrass oil is obtained by methods such as steam distillation. In addition. the processes produce only small amounts of oil. the pectinase application. lemongrass oil is also used as starting materials for Vitamin A. quality and stability. thus. sprays and perfume preparation. Extraction by volatile solvent. To compare the physico-chemical properties of the control and enzymetreated samples. Steam distillation and solvent extraction are used commercially. 1997). lemongrass is a very important oil component in soaps. It is enery-efficient and economically feasible (Espino. 1997).
Laguna from May 1-31. 1982). Significance of the Study The Philippines is dependent on importance of essential oils from Europe and the United States. quality And stability of Lemongrass oil Fig. Our country practically imports more than 90% of our requirements and ranks 8th among the essential oil importing countries (Dar. This project can be considered as an initial phase to scale up the production of essential oils using the pectic enzymes previously established by the BIOTECH. 1. Los Baños. 1999. thus. Laboratory from August 28. The researcher was concerned in determining the percent essential oil yield and in performing its physic-chemical analysis. 1997). . The second trial was conducted at Cinco’s Residence and the Filipinas Palm oil Plantations.December 29. alleviating the supply of essence and coping with the increasing demand. The traditional method of essential oil extraction is by steam distillation. 1999. The study focuses only on the effect of the enzyme pectinase on the production of essential oil from lemongrass. better quality and stability.1 Paradigm of the Independent and Dependent Variables of the Study “Pectinase-enhanced Production of Essential Oil from Lemongrass (Cymbopogon citratus)” Scope and Limitations of the Study The study was conducted at the Immunology and Enzyme Laboratory of the National Institute of Molecular Biology and Biotechnology of the University of the Philippines. This study was conducted to develop a new technology in extracting essential oils from locally available lemongrass that will account for maximum yield. despite the fact it is one of the tropical countries rich in essential oil-bearing plants (Anzaldo. The researchers also utilized the available laboratory apparatus such as the Electromantel oil extraction evaporator equipment of the FPPI palm oil mill laboratory. It can be the basis for other pectinase applications. the process involves the use of much energy and it yields small quality of oil apart from loosing some of its components. Inc. However.Theoretical/Conceptual Framework INDEPENDENT VARIABLES Control (without enzyme) Treatment 1 (w/ enzyme) DEPENDENT VARIABLES The yield. Data gathering and statistical analysis has the limitation of bias.
. useful in many industrial processes. . This is also in support to the long standing concerns of economic stagnation in the rural areas of many developing countries by the United Nations.a substance capable of maintaining the relative concentrations of hydrogen and hydroxyl ions in a solution by neutralizing.combination of organic acids and alcohol. it is an “agro-based industry that can utilize rural sector participation in the cultivation and harvesting of raw materials and to some extent. capable of maintaining the pH of a solution.” The improvement of the agro-techniques and the development of energyefficient and affordable technology in the extraction process can add value to the product and increase its market potential.an unsaturated liquid aldehyde C9H15CHO that has a strong lemon and verbena odor. . It can also be a strategy in solving the importation problems of foreign made essential oils. As it is economically feasible. As stressed by UNIDO. . a large scale production for commercial purposes will soon be possible in the country.The use of pectinase has several advantages in contributing to a more economical process.any of a very large class of complex proteinaceous substances that are produced by living cells that are essential to life by acting like catalyst in promoting chemical reactions in the cell without being used up. sweet – smelling compounds widely used as artificial flavors and dye.any of five isometric volatile liquid paraffin hydrocarbons found in petroleum especially the normal hydrocarbon.product of the reaction of two molecules of alcohol. used in flavoring and in perfumery and consists of a mixture of two stereoisometric forms. Buffer . Definition of Key Terms Aldehyde . The program for the development of the essential oil industry was perceived to answer these concerns.production by the oxidation of alcohol compound containing the CHO group and used in the manufacture of dye and synthetic rubber. Citral Enzyme Esters Ethers Hexane . . very volatile and highly flammable. even in field of distillation activities.
Espino et al. also studied the pectic enzymes of A. the cost per ml and cost per pectin transeliminase (PTE) unit of crude pectinase was PO. They can hamper processing or lower the quality of a product. are of major importance to food industry. Pectic enzymes ca also be utilized in the pre-treatment of pectic wastewater. 1984). Espino at BIOTECH – UPLB. macerate vegetables and other applications (Godfrey.034 respectively (Espino. Pectinase was isolated from Aspergillus niger grown by solid substrate cultivation in enameled trays placed in cabinet type fermentor. Today. 1986). are used for commercial production of these enzymes. Molds especially the species belonging to the Aspergillus family.. During stepwise extraction of pineapple juice. pectic enzymes causes these substances to easily degrade. using pectic enzymes.M. . The enzyme (pectinase) used in the study was provided by the group of Dr. 1983. 087 and P 0. Another important industrial application of pectinases is in the degumming of fibers crop like ramie (Deshpande and Gurrucharanan. Pectic substances are found in the wall. guava. Isolated from various substrates. (1989) partially isolated pectic enzyme extracellularly-produced by Aspergillus ficuum and A. or papaya without addition of too much water that affects the characteristics of fruit flavor and aroma. pretreat cereals. intercellular spaces.CHAPTER 2 REVIEW OF RELATED LITERATURE The Pectic Enzyme and Aspergillus Family Pectinases are produced by molds. The enzyme produced high pectolytic and clarifying activities. it is possible to extract juice fruits from fruits such as banana. accelerates rotting of flax. enzyme addition also improves the quality of juice by allowing extraction of more soluble solids and juice particles. 1983). They are utilized in the fruit juice industry to facilitate extraction and clarification of juice (Forgat and Kelly. Espino et al. Whitaker. Several studies have been done describing the purification and properties of pectic enzymes produced by Aspergillus sp. 1972). niger isolated from pomelo (Citrus grandis Osbeck). remove mucilage in coffee after fermentation. 1997). these enzymes have been adopted to improve the yield of alginate base materials from harvested seaweeds. yeasts and bacteria. Economic analysis of pectinase production showed that. Pectic enzymes are also used for maceration and liquidation of fruits and vegetables. ficuum isolated from local agricultural wastes. Studies on Pectinase Application Pectic enzymes. T. beans and pulses to reduce cooking times and temperatures. 1985). hence. juices and saps of higher plants (Whitaker. (Tanabe et al. In the industrial processing of plant tissues. which degrade pectic substances.
valued at US$ 700 M. Department of Trade and Industry). we have been exporting oils of peppermint. Results of the experiment showed that there was an increase of about 75% in the yield of the enzymetreated sample and the quality was considered similar to that of the extra grade oil while the untreated sample are of lower quality essential oils. United States of America. (4) aldehyde. (5) ketone. 1993). The Philippines is one of the principal countries that is very dependent on imports from China. Essential oils vary from colorless to yellow or brown and are usually a mixture of compounds. Our net import of these commodities amounted to US$ 58 M for 1993 (Jamilla. Their almost complete volatility in steam distinguishes them from so called “fixed” or “fatty” oils such as coconut oil or palm oil. Verlet (1993) estimated that world production of essential oils is around 45. (Espino et al. which collectively are responsible fro imparting the characteristics odor associated with the plant material itself (Wijieskera and Tatnatunga. The importance of essential oils can be gleaned from their extensive utilization in various industries. accompanied by the building up of tariffs and quotas on importation. commercial production of these oils. All these figures combined show our tremendous dependence on importation of our essential oil needs. The Essential Oils Essential oils are volatile constituents of plant material. Our export of essential oils declined thereafter but rose again in 1993.To increase the oil yield and improve its quality. Based on the data. (8) acids and (9) terpenes. oxidation. An increase of 8. India and Europe. lemon. which are derivatives of glyceric acid.000 tons.. (6) lactones. but these commodities are most likely re-export materials as we do not have any local. Montedro et al. 1997). etc. (Based on the statistical data obtained from the Foreign Trade Statistics. 97) phenol. They are also sometimes called “volatile” or “essential oils”. turbidity. (3) esters. induction time.9% in olive oil yield and generally improved quality. Current Market of Essentials Oils. The compounds occurring in essential oils are as follows: (1) hydrocarbon. chlorophyll and contents of aromatic compounds resulted from the study. spearmint. the essential oils are mixtures of a variety of chemical ranging in number from few to several. The UPLB BIOTECH – produced pectinase was also used for the extraction of essential oils from ilang-ilang flowers. They serve as major and minor constituents in the manufacture of perfume and cosmetics. (1993) investigated on the pectinase from olive oil vegetation waters and its use in the mechanical olive oil extraction process. (2) alcohol. Indonesia. The highest value and amount of export was observed n 1989. In 1989-1993 there is an increasing trend in the value of importation of essential oils. Import substitution in the traditional sense of replacing imported essential oil with domestic sources of production and supply. 1997). Brazil. is likely to be considered by our policy- . flavoring agents in food and confections and pharmaceuticals for treatment of ailments.
Aldehyde content contributes to the aging of essential oils. Solvent extraction is technically a more advanced process and yields real representative odors. Extraction of Essentials Oils. It has hypotensive property which lowers blood pressure when used in the experimental animals.8876 to 0. as they may or may not be the same components.. 1984). The low priority that has been given to the development of essential oils industry is what makes the development more challenging. Distilled oils may also be considered artifacts. but is more expensive than distillation. Ionones are now employed for the manufacture of synthetic Vitamin A. It is effective as a liniment for chronic rheumatism. etc.14 mg/g: and citral content by GC = 75% minimum. mythyl ionone. Lemongrass. Essential oils are usually recovered from plants by two methods. It is widely used as flavorant for food and pharmaceutical industries. As the Dean of the College of Agriculture said “there was money in the essential oils business for even hunger was no excuse for a person not to smell good. (2) specific gravity at 20oC = 0. Steam distillation. either by distillation or by solvent extraction. It is not only known for its essential oil but also for the medicinal value of the palnt itself (Coronel et al. It is therefore advisable to engage in new technologies and techniques in the extraction of essential oils that would be cheaper and increase the oils yield and stability. which are lost during the process. The physio-chemical properties of lemongrass oil based on the study conducted by Torres (1994) are as follows: (1) refractive index at 20oC = 1. on the other hand. The Lemongrass Oil Literature about the nature of essential oil found in lemongrass is very limited due to lack of knowledge that it can be a substitute source of essential oil. locally known as “tanglad”. During the process.4892. as well as for chemical transformation into ionone. limbago.makers to be out of tune with the trade globalization and liberation measures that have been adopted by the government (Jamilla. the oil contains at least 75% citral (Bellanato and Hidalgo. 1984). nueralga. contains oil having a strong. sprains and other painful affections and ringworm (Quisumbing.04 to 14.9215. . the collected mixture may consist largely of steam with only small portion oil. lemon odor due to its citral content. 1997). This is the real challenge that undoubtedly calls for a concerted multidisciplinary approach. (3) solubility in ethanol (v/v in 70%)=1:1. (4) acid value = 11. The oil is useful in flatulent and spasmodic affections of bowels and in gastric irritability. Lemongrass oil is a main source of citral for lemony and verbena notes. 1978). is a very much more gentle process and causes few drastic changes in the oil.
.5% of the sample 12 hours 28oC 1:6 Incubation Time Incubation Temperature Sample: buffer dilution Ratio Source of Materials 12 hours 28oC 1:6 Freshly harvested lemongrass were obtained from Los Baños and Quezon (for trials 1. Laguna. University of the Philippines Los Baños College. 3. The lemongrasses were cut into pieces and ground in an osterizer. Espino of the National Institute of Molecular Biology and Biotechnology (BIOTECH). Table 3. Agusan del Sur (for trials 4 to 8).CHAPTER 3 METHODOLY The Research Design The experimental design used in this study was a two-group design wherein a control and a single treatment were manipulated (Table 3.60 PTE units 0.1 Research Design Composed of the Control and the Treatment CONDITIONS Lemongrass Enzyme (pectinase) CONTROL 500 g absent TREATMENT 500g 2. The average pectinase transelinmase activity of the enzyme used in this study was 24.5 g (61. 2 and 3) and Purok 3 Brgy.64 units per gram of ammonium sulfate precipitated pectinase. The pectic enzyme used in this study was provided by the group of Dr.1) there were eight trials conducted both in the control and the treatment. Enzyme Treatment Freshly harvested lemongrasses were processed immediately for extraction. San Francisco.
Extraction Procedure The reaction mixture composed of the sample. Twenty panelists were invited to evaluate the odor rate of the oil. Lastly. pH 4. The extracted essential oil is then filtered using filter paper. For trials 4 to 8 the solvent was recovered from the essential oil by the use of an Eloctromantel oil extraction evaporator equipment in the oil mill laboratory of Filipinas Palm oil Plantations. The flask was connected to the rotary evaporator system to separate/recover the solvent from the essential oil. In measuring the percent yield of the lemongrass oil. both in the control and treatment. & 3). for maximum market production of lemongrass oil. the researcher used the available analytical balance. After incubation.The samples were weighed and then the enzyme solution was added. the reaction mixture was cooled to room temperature before extraction. the reaction was stopped by placing the mixture in a boiling waterbath for 10 minutes to inactive the enzyme. The researcher also subjected the sample to physic-chemical analysis. enzyme solution and solvent were filtered using cheesecloth. The results were compared to the literature value as the standard basis for comparison. The panelists were given score sheets for rating the odor using the 5-point rating scale. The extract was transferred into a weighed flask. However. A control consisting of the ground lemongrass leaves and buffer was also prepared. The panelists were chosen for their knowledge on essences. The reaction mixture was incubated at 28oC with constant shaking for 12 hours. . Manual squeezing was done to recover most of the liquid. Data Gathering Technique After the extraction process the essential oil recovered from lemongrass was weighed.2 M acetate buffer. were placed in a single room for the panelists to evaluate. The percent yield of the oil was the determined using the formula: Percent essential oil yield = weight of essential oil x 100 weight of the plant sample The process was used from the second to the eight trials both in the control and enzyme-treated samples. The process was stopped when the content of the sample flask was perceived viscous (for Trials 1. Samples for each trial. The enzyme solution was prepared by dissolving the enzyme in 0. 2.0 at !:6 sample: buffer dilution ratio. the use of machine operation is highly recommended. The filtrate was collected while the residues were discarded. Inc. The reaction flask was sealed tightly to prevent any loss of essential oils.
The values obtained were compared with those reported in the literature. Arithmetic mean. . t-test and analysis of variance (ANNOVA) were the statistical tools used in analyzing and interpreting the data. ANNOVA and t-test were used to determine the significant difference of the results. 2. The extracted essential oil from lemongrass was analyzed as to its specific gravity and solubility. Quality analysis of the essential oil. Comparison of essential oil yields between the untreated and enzymetreated samples.Data Gathering Method The results were gathered and tabulated. Mean was used to determine the average of the percent yield of the oil as well as its rate. Quantitative and qualitative analysis were done based on the following: 1.
5% of the sample + 3L. 0. to inactive the enzyme Cooling to room temperature Filtering with cheesecloth Extracting with petroleum ether as solvent Recovery of organic layer in a separatory funnel Recovery of essential oil by rotary evaporation or Electromantel oil extraction evaporator Filtering with filter paper Weighing of recovered oil and determining the percent yield Physical and chemical analyses of the recovered essential oil .0) Processing of Lemongrass (cutting into pieces and grinding using an osterizer) Addition of enzyme solution (1:6 sample: buffer dilution ratio) Incubation of reaction mixture at room temperature For 12 hours with constant shaking Boiling in waterbah for 10 mins. pH 4.2 M buffer.FLOW CHART OF THE PROCEDURE Enzyme preparation (enzyme (2.50g) 0.
1) were obtained.treated lemongrass belongs to the extra grade oil category.96%) exceeded the literature range value of 0.78% of the total weight of the sample.48% to 0.like odor. application of pectinase on essential oil extraction truly enhances and increases the essential oil yield. the specific gravity of the enzyme. Moreover. The use of pectinase showed that enzyme treatment5 improved the quality of lemongrass oil. Results of the analyses showed that the enzyme treated lemongrass oil had higher specific gravity (1. Table 2 presents the physicochemical properties of the extracted essential oil lemongrass.60 PTE units) at a sample to buffer dilution ratio of 1:6 (w/v). essential oil extraction from lemongrass is feasible.987**). Physico-chemical Analysis of the Essential Oil from Lemongrass The extracted essential oil was a golden yellow colored liquid (deepest color base on the color range indicated in the literature) and has a lemon.2% to 0.033) than the control (without enzyme) with only 0. The use on enzymatic process could be the key to provide the growing demand of essential oil in the industry today. 61. ANALYSIS AND INTERPRETATION OF DATA The results of this study were shown in the preceding tables and figures. In addition to these. Based on the results obtained. There is a significant difference on the average percent essential oil yield between the control and the enzyme-treated (2.47% for the control and 0. 1983).96% for the enzyme treated samples (Table 4. other physical and chemical properties were also examined and compared. the enzyme treated oil appeared to be darker than the oil obtained from the control sample.5% of the sample. Results showed an essential oil percent yield of 0. The specific gravity is directly related to the quality of the oil. 1997).Chapter 4 RESULTS. . However. Large-scale extraction of essential oil from lemongrass could be explored and feasibility studies on economic viability of the process in order to attract investors. the characteristic fragrance of lemongrass. under 280C and for 12 hours incubation time.4% can be considered normal (Brandares. the oil yield obtained in this study for the enzyme-treated samples (0. with the use of a minimum amount of enzyme (0. Based on the literature value (Robbin. Yield of 0. Based on the physical appearance. The oil yield of the enzyme-treated sample was approximately twice than that of the control (without enzyme). 1987). Furthermore.676. Comparison of Essential Oil Yields between the Control and Enzyme treated Samples Previously established optimum conditions were adopted in extracting essential oil from 500g lemongrass (Galang. the computed t-value exceeded the tabular F-value in the 5% and 1% probability.
the solubility in 70% ethanol (v/v) of the enzyme-treated lemongrass oil is the same as that of the literature value.63 0.05(8) = 1.25(8) = 2.42 0. Therefore.39 0.95 0. On the other hand. enzyme treatment resulted to better quality essential oil as compared to the control.96 0.64 0. Table 4.63 0.1 Comparison of the Percentage Essential Oil Yield of the Control and Enzyme-Treated Lemongrass Trial 1 0.5% enzyme concentration 12 hours incubation time 280C incubation temperature 1.6 sample: buffer dilution ratio : : 2.1(8) = 2.47 0.99 0.0306 t 0.94 Trial 7 0.860 t 0. At the same time.896 T-test values: Computed t-value Tabular t-value . Based on the results of the physical and chemical analyses.96 Treatmen t Control Enzymetreated Average 0.98 Trial 2 0.38 0. the increase in essential oil yield as a result of enzyme treatment compensated the cost of the enzyme added.41 0. As perceived by the 20 panelist. the enzyme-treated lemongrass oil has a higher odor rate compared to that of the control. it can be concluded that pectic enzyme treatment under optimized conditions at laboratory scale can be an alternative method for essential oil extraction of lemongrass on large scale.the value is a little bit lower compared to the commercial oil.23 0.987** t 0.98 Trial 8 0.96 Conditions used: 0.94 PERCENT YIELD Trial Trial Trial Trial 3 4 5 6 0.
1 (v/v). Moreover.5%. Results on the physical and the chemical analyses of the extracted essential oil showed that enzyme treatment improved the quality of the oil compared to the control (without enzyme). sample: buffer ratio 1:6 (w/v). Therefore. An essential oil yield of 0. the application of peptic enzyme at laboratory scale could be an alternative method for essential oil extraction on a large scale. incubation temperature: 280C. . there is also a significant difference on the odor rate of the lemongrass oil as perceived by the 20 panelists. On the other hand.47% oil yield for the control (without enzyme). Conclusion The application of the pectinase in the extraction of essential oil from lemongrass was found effective. incubation time: 12 hours. the use of enzymatic treatment contributes to a more economical process and is therefore economically viable. Moreover.96% was obtained for the enzyme-treated sample while 0. Thus. there is a significant difference on the mean of the control and of the enzyme-treated samples. the researcher believed that the enzyme treatment increased the essential oil yield and further improved its quality and stability. This shows that pectinase application doubles the yield of the extracted essential oil from the lemongrass. these properties were comparable with those reported in the literature. The pysico-chemical properties of the enzyme-treated lemongrass oil were as follows: specific gravity = 1.Chapter 5 SUMMARY OF FINDINGS. CONCLUSIONS AND RECOMMENDATIONS Summary of Findings Essential oil from lemongrass was extracted using the pectinase under the following optimized conditions: enzyme concentration: 0.033 and solubility in 70% ethanol = 1.
and 3.Large-scale extraction of oil from lemongrass through enzymatic process should be further explored and feasibility studies on the economic viability of the process should be conducted. . 2.Characterization of lemongrass essential oil components should be made in order to determine which is responsible for the characteristics of lemongrass odor. the researcher recommends the adaptation of this new technology on the extraction of essential oils. the researcher also recommends the following: 1. With this basis . Furthermore.Recommendations Production of essential oil from lemongrass through enzymatic process yielded positive results. application studies on other oil-bearing plants apart from lemongrass should be explored.Other physical and chemical analyses should be conducted on the extracted essential oil to further evaluate its quality. To further improve the study.
Storage of maize and cowpea and inhibition of microbial agents of biodeterioration and biodegradation. De Guzman. Espino. 1987. Prospects of Essential Oil Business in the Philippines. D.Q.June 1994. 1997. J.O. V. Application of Pectinase on the Extraction of Essentials Oils from Lemongrass (Cymbopogon Citratus). Paper presented to the 1st SARCENHP in June 1997.P.. M. et al 1993. 290-291.Q.A. T. of food Science 5 (4):355-362. V. . Phil. Heyden and Son Ltd. et al.E.. NSTA Technology Journal IX (3):36-28.R. F. W. Ricalde and F. Jou. July 1993. Annual Report. Pectinase production from olive vegetation waters and its use in the mechanical extraction to process increase oil yield and improve quality. Begliomini. Horticulture Production of Essential Oil Plants in the Philippines Paper presented in the 1st Southeast Asian regional Conference and Exhibition on Natural and Herbal Products (SARCRNHP) in June 1997. Ltd. Montedro.M. II Cymbopogon citrates (DC) Stapf. J. June 1995.B. A new development in cologne formulation using ilang-ilang flower oil. Darjuan. and B. Coronel and F. 2nd ed. A. Sanchez. Jamilla. 1999. Coronel. G. Forgarty. and C. NSTA Technology. B. A. England. Manalo.M. B. Elsevier Science Publishers. Kelly. 7 (2): 21-34.B. Bellenato.M. G. Galang. 6(2): pp. Stability Studies of Essential oils from Philippine Plants. Sci 116 (4):391-402. Annual Report on Essential Oil Research and Development Program (DOST-PCCARD). C. Vuelban. 1984.C. Pilot Production and Utilization of Peptic Enzymes from Food Applications. Odesola. Infrared Analysis of Essentials Oils. Effect of moisture content on the essential oil yield of lemongrass and citronella. 81-84.T. Guzman. p.. P. pp. Anzaldo. J. C.C. Italian Journ..F.E.E. Anzaldo and M. M. 1035 Brandares. 1982. 1990.L. Madrid. Microbial Enzyme and Biotechnology.F.T. M.Nov. 1994. and Hidalgo. Anzaldo. Recaña.References Adegoke.
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