Experimental Hypothesis The experimental hypothesis is that the process of steam distillation will yield a higher percent recovery of Eugenol

from cloves and will have fewer impurities than the vacuum adaptor extraction process using the Eugenol mash.

Abstract Results

Future Research

Materials and Methods

Natural Product Isolation and Extraction of Essential Oils

Jared A. Taylor (J. Abrams Advisor) Bluefield State College Bluefield, WV

Acknowledgments This research was funded by Bluefield State College, advised by Dr. Jason Abrams, and assisted by Andy Terry .

Eugenol is the primary compound that is found in cloves and is also known as clove oil. The medical field uses Eugenol quite frequently as an analgesic, antiseptic, or even as a local anesthetic. The experimental hypothesis is that the process of steam distillation will yield a higher percent recovery of Eugenol from cloves and will contain fewer impurities than the Eugenol mash employing vacuum adaptor filtration. The steam distillation of essential oils from cloves was performed and Eugenol was collected. A higher percent recovery increases the value of the natural product.

Purity Profile: The composition of the Eugenol extract was a) analyzed on Thin Layer Chromatography by employment of a 254 nm UV Lamp and Retention factor (Rf) values were-calculated. b) Three staining techniques (Vanillin, Iodine Chamber, and Potassium Permanganate Stain) were also performed to test for components that were not UV active. Yield Profile: The percent recovery was 5 % or 0.4 grams from the 8 grams of original ground up cloves. The Vacuum adaptor extraction of the Eugenol mash was performed and a 3.4 % or 0.136 grams recovery was obtained from the Eugenol mash.

For Eugenol, exclusively: Our findings showed that an average steam distillation Rf = 0.76 (Ethyl acetate –Hexane mixture (3:7) compared to average vacuum extraction with Eugenol mash had Rf = 0.71 (Ethyl Acetate –Hexane mixture (3:7). This indicates that the Eugenol obtained from the steam distillation achieved a greater purity than that collected from the vacuum extraction.

Eugenol is the primary compound that is found in cloves and is also known as clove oil. The medical field uses Eugenol quite frequently as an analgesic, antiseptic, or even as a local anesthetic. The experimental hypothesis is that the process of steam distillation will yield a higher percent recovery of Eugenol from cloves and will contain fewer impurities than the Eugenol mash employing vacuum adaptor filtration. The steam distillation of essential oils from cloves was performed and Eugenol was collected. A higher percent recovery increases the value of the natural product.

Purity Profile: The composition of the Eugenol extract was a) analyzed on Thin Layer Chromatography by employment of a 254 nm UV Lamp and Retention factor (Rf) values were-calculated. b) Three staining techniques (Vanillin, Iodine Chamber, and Potassium Permanganate Stain) were also performed to test for components that were not UV active. Yield Profile: The percent recovery was 5 % or 0.4 grams from the 8 grams of original ground up cloves. The Vacuum adaptor extraction of the Eugenol mash was performed and a 3.4 % or 0.136 grams recovery was obtained from the Eugenol mash.

For Eugenol, exclusively: Our findings showed that an average steam distillation Rf = 0.76 (Ethyl acetate –Hexane mixture (3:7) compared to average vacuum extraction with Eugenol mash had Rf = 0.71 (Ethyl Acetate –Hexane mixture (3:7). This indicates that the Eugenol obtained from the steam distillation achieved a greater purity than that collected from the vacuum extraction.

Eugenol is a compound that is found in cloves, and because of this it is sometimes known as clove oil. The chemical formula is C10H12O2 It has a very distinct smell and is used in some perfumes. Eugenol is soluble in organic solvents and its appearance can be a soft yellow or even a clear oily liquid. The medical field uses Eugenol quite frequently as an analgesic, antiseptic, or even as a local anesthetic7. It can help with pain during dental visits and also act as a numbing agent if a patient gets “dry sockets” after a tooth extraction. Natural product extraction is well utilized in food, medical, and manufacturing industries. The biosynthesis of eugenol is highlighted below.

The extraction process is only one step in the isolation of pure compounds from plants

The next step in this experiment would be the process of Flash Chromatography to separate and test the impurities in the different extraction processes, such as Eugenol acetate and sesquiterpene. In this process, non-polar compounds will elute from the column first before the polar compounds. Acid-Base extraction process could also be used to separate Eugenol acetate by using sodium bicarbonate. The procedure of acid-base extractions uses serial liquid-liquid extractions to purify the acids and bases from the mixture.

Other methods must also be utilized in order to further purify natural products, including the use of recrystallization, distillation, and column chromatography. The extraction process encompasses the use of a variety of organic solvents with different solubility for substrates and due in part to differences in solvent polarities. Accompanying the extraction process is the employment of thin layer chromatography (TLC) in order to assess the purity level of materials once the extraction process is complete, and to determine what course of purification must be done next (additional extractions, column chromatography, distillation, or recrystallization).

Steam Distillation Vacuum Adaptor Extraction

to vacuum

filtrate contains essential oils

insoluble lignans filtered off

bed of celite

Crude Products to Be Analyzed

Steam Distillation is an efficient method of extraction that is used to separate the essential oils from cloves. The steam distillation involves distilling a mix of ground up cloves and water to obtain the oils. The 0.8 grams of cloves that were used in this experiment are ground up using a mortar and pestle, in order to increase the surface area, and added to a 250 ml distilling pot with 100 ml of water. This process uses a 3-way connector to attach the straight condenser, which is attached on the other side to the receiving pot or flask.

TLC Stains

Vacuum Extraction using a Eugenol mash was the second extraction process in this experiment. Using 4 grams of ground up cloves by using a mortar and pestle and 50 ml ethyl acetate created a Eugenol mash.

UV Active Spots

Thin Layer Chromatography (TLC) was then used to assess the purity of the Eugenol recovered from the steam distillation against the vacuum extraction using Eugenol mash. The TLC is composed of a polar stationary phase, silica gel, with an alumina backing. Eugenol is seen most clearly as the blueish-brown spot in the KMnO4 stained plate.

THIN LAYER CHROMATOGRAPHYTHIN LAYER CHROMATOGRAPHY

The steam distillation of essential oils from cloves was performed and Eugenol was collected. The percent recovery was 5.0 % or 0.4 grams from the 8.0 grams of original ground up cloves. The Vacuum adaptor extraction of the Eugenol mash was performed and a 3.4 % or 0.136 grams recovery was obtained. The samples of Eugenol were then tested on Thin Layer Chromatography and Retention factor (Rf) values were obtained for comparison of purity from each sample of Eugenol. Average steam distillation Rf = 0.76 (Ethyl acetate –Hexane mixture (3:7) compared to average vacuum extraction with Eugenol mash had Rf = 0.71 (Ethyl Acetate –Hexane mixture (3:7)

SummaryThe hypothesis that the steam distillation would have fewer impurities and yield a higher percent recovery was supported by our experiments. In conclusion, the steam distillation is a more efficient method of extracting Eugenol from cloves, and should continue to be used commercially to achieve economy of scale. The mash procedure was not as selective as the steam distillation. The 5.0 % recovery from the steam distillation process is similar and comparable to the literature value for this experiment. A higher yield/recovery value increases the value of the natural product.

O OH

NH2

OH

H3CO

OH

H3CO

OH