The lutein molecule

Chemical structure from Lutein Information Bureau, 2006.

The Concentration of Lutein in Ocimum basilicum

Regan Feuerhelm 8th grade Lane School Lane, OK 74555

A Botany Paper Presented to Oklahoma Junior Academy of Science February 21, 2009

Abstract This experiment was to determine which of the four varieties of basil, Genovese, Nufar, Ethiopian, and Mrs. Lemon Burns, grown in an organic field and a conventional field, contains higher concentrations of Lutein. However, if organic production methods decrease the basil flavor then people will not buy the herb. A small portion of fresh Genovese basil was used for taste tests to see if people preferred organic or conventionally grown basil for taste, color, texture, etc. All four varieties of basil from both fields were harvested on the same day, stored at -80OC, and on a later date, the lutein was extracted with water and ethanol. Samples were put in a glass cuvette and read with a spectrophotometer where the computer takes transmitted light and converts it to an absorbance reading. This reading determines the amount of lutein in each sample. The data shows that Ethiopian has the highest lutein concentration. Also conventional basils have higher lutein concentrations than organically grown basil. Taste test scores showed little or no differences between the taste, texture, color, or aroma of conventional or organically grown basil which showed that organically produced basil will not ruin the taste and people will buy it.

Introduction Ocimum basilicum, more commonly known as Basil, is an herb commonly used to liven up the flavor in about any dish (Basil, 2009), but can it help prevent Age-related Macular Degeneration (AMD), a disease that deteriorates pigmentation in the eyes. Previous studies have shown that basil is a good source of lutein (Basil, 2009), which is associated with preventing AMD (Richer, S., 2004; Lutein, 2009). It is suggested to eat 6 mg of lutein each day (Lutein Information Bureau, 2006). This experiment is to determine which of the four varieties of basil, Genovese, Nufar, Ethiopian, and Mrs. Lemon Burns, grown in an organic field and a conventional field has a higher concentration of Lutein, an antioxidant that helps prevent diseases like AMD. However, the consumer may not purchase the basil if it doesnt have an appealing taste, color, or texture; so during this experiment two taste tests on organic and conventional Genovese, the typical pesto basil, were preformed to test the acceptability of the basil flavor. Does organic food taste better? No research has shown that organic food tastes better than non organic food. However, people strongly feel that organic food tastes much better than non organic food (RuralTech Services, 2006). A previous study linked lutein concentration to total chlorophyll concentration (Nishiyama et al., 2005). So the darker varieties should have more lutein. Since Genovese was the greenest variety, it should have the most lutein. And since the organic leaves seemed greener, organic basil should have more lutein. Because of these initial observations, the following two hypotheses were developed. 1) Since lutein amounts

correlates with chlorophyll concentrations, the hypothesis in this experiment is that Genovese will contain a stronger concentration of lutein than the other basils tested, because it had a darker green leaf. 2) The second hypothesis is that organic basil will contain more lutein and have a better taste, texture, color, and aroma than the conventional basil because it does not have the pesticides used on conventional basil.

Methods This experiment started in the summer at the South Central Agriculture Research Center where four varieties of basil were harvested and then frozen for future testing. Then a small portion of fresh Genovese basil was used to prepare the pesto (Simple Recipes, 2009) and marinara pasta taste tests to see if organic or conventionally grown basil has a better taste, color, texture, etc. The people in this experiment rated the basil between one being the least desirable and four being the most. Then the scores were averaged to start on the graphs. After the taste tests all varieties of basil from both fields were taken from the freezer and the leaves were plucked from the stem and weighed. Ten grams of each were used. It was then pulverized with a mortal and pestle and mixed and mixed in a solution of 10 milliliters ethanol and 30 milliliters water. Then the solutions were put into vials with caps and then was put in a centrifuge that spins at 10,000 cycles a minute so the liquid goes to the top and the basil leaves at the bottom. The lutein is extracted into the liquid (Craft, N. 2001). The samples were then put in a glass cuvette. Each cuvette was than put in a spectrophotometer where the intensity of the wavelengths are measured to determine the

amount of lutein in each sample. This is done when a bright light shines through the glass and a detector on the other side picks up the green and yellow wavelengths from the liquid substance. Then the computer takes transmitted light reading and converts it to an absorbance reading (Microsoft Encarta Online Encyclopedia, 2004). The equation to convert absorbance readings to lutein concentration was [x g/ml lutein=(1000(A470 2.13 (Ca)- 97.64(Cb))/209]. Graphs were made using Microsoft Excel.

Results and Discussion Lutein concentration in four variety of basil. Once it is all tabulated the data shows that Ethiopian has a higher concentrations of lutein over all the other basils. The averaged Ethiopian samples had 189 g/ml lutein. Lemon and Genovese were almost tied for a median range of lutein 182 and 179 respectively. Nufar had the least at 163 g/ml lutein. Taking into account our dilution factor, Genovese had 716 g lutein /g fresh basil leaf. If your only source of lutein was basil, to get the daily amount of lutein suggested by the Lutein Information Bureau, you would need to eat 8.4 grams basil every day. Rejecting the hypothesis that Genovese had more lutein. Genovese appeared to be darker green, but because Ethiopian has a purple tint to it, the purple might have hidden some of the green color. As a side note, we noted for future experimentation that it would be better to Add the 10mL ethanol and then the 30mL of water instead of adding a water ethanol mixture. This would result in a clearer reading (less haze) in the spectrophotometer.

Lutein concentration in conventional vs. organically grown basil. The data shows that the conventionally grown basil has a stronger concentration of lutein than the organically grown basil. An average of 212 g/ml lutein in the conventional basil and 144 g/ml lutein in the organic basil. The hypothesis was rejected because organically grown basil had less lutein conventionally grown basil. One possibility is that since the conventionally grown plants had much smaller leaves which might have caused a concentration of the lutein. Originally it was assumed that the green color would associate with lutein concentration did not hold up in our studies. However, it is possible that visual ratings of how green a leaf is subjective. A light green might look more green to some, but a dark green would have more chlorophyll.

Taste test. The result of the taste tests showed that there is little or no difference in the texture, aroma, color, taste, or basil flavor between conventionally and organically grown Genovese basil. That means that people will still buy the basil whether grown conventionally or organically. Once the taste tests scores were averaged it showed that there was little or no differences between the taste, texture, color, or aroma of the conventional or the organically grow basil.

Conclusions In conclusion the data shows that Ethiopian has a better overall average of lutein then the other varieties of basil. Also the data shows that the conventionally grown basil has a stronger concentration of lutein than the organically grown basil. The result of the taste tests showed that there is little or no difference in the texture, aroma, color, taste, or basil flavor.

Acknowledgments I would like to thank PhD Angela Davis for helping me with my experiment and giving me the suggestion for this experiment. I also would like to thank Amy Helms for helping me pulverize the basil leaves for the testing for lutein concentration. I would especially like to thank Mikki Runyan for allowing me to perform this experiment. I thank Amy Helms for taking my picture.

Bibliography 1.) Microsoft Encarta Online Encyclopedia. 2004. [on-line]. Available from http://www.spectrophotometer.com/FAQ.htm. (Accessed Feb. 21, 2009). 2.) Basil. 2009. [on-line]. Available from http://en.wikipedia.org/wiki/Basil. (Accessed Feb. 21, 2009). 3.) Lutein. 2009. [on-line]. Available from http://en.wikipedia.org/wika/Lutein. (Accessed Feb. 21, 2009). 4.) Lutein Information Bureau. [on-line]. 2006. Available from http://www.luteininfo.com/about. (Accessed Feb. 21, 2009). 5.) Simple Recipes. 2009. [on-line]. Available from http://www.elise.com/recipes/archives/001329print.php. (Accessed Feb. 21, 2009). 6.) RuralTech Services. 2006. [on-line]. Available from http://www.organicfacts.net/organic-food/organic-food-basics/organic-foodfaqs.html. (Accessed Feb. 21, 2009). 7.)Nishiyama, T. Fukuda, T. Oota. 2005. CULTIVAR DIFFERENCE IN CHLOROPHYLL, LUTEIN AND -CAROTENE CONTENT IN THE FRUIT OF KIWIFRUIT AND OTHER ACTINIDIA SPECIES. Acta Hort. (ISHS) 753:473-478. Craft, Neal, E. 2001. Protocols In Food Analytical Chemistry. (.E. Wrolstad, et al. eds). p. F.2.2.3. 8.) Richer, S., W. Stilies, L. Statkute, J. Pulido, J. Frankowski, D. Rudy, K. Pei, M. Tsipursky, J. Nyland. Double-masked, placebo-controlled, randomized trial of lutein and antioxidant supplementation in the intervention of atrophic age-related macular degeneration: the Veterans LAST study (Lutein Antioxidant Supplementation Trial). Optometry, 2004. 75(4): p. 216-30.