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Technical Literature Kemin Health, L.C. 600 East Court Ave, Suite A Des Moines, Iowa 50309-2021 USA tel: 515.248.4000 fax: 515.248.4051 toll free: 888.248.5040 www.kemin.com Prevalence of Versus Esters in Human Diets Zoraida DeFreitas, Ph.D. Director, Research and Development Key Conclusions 93% of the total amount of lutein, zeaxanthin and their found in human diet are present as lutein and zeaxanthin and not as. and zeaxanthin, as found in marigold extract, are not the predominant sources of lutein and zeaxanthin found in a typical diet. Introduction Everyday people are discovering the benefits of including important antioxidants in their diets to maintain better health. In particular, people concerned about Age-related Macular Degeneration (AMD), the leading cause of blindness in the industrialized world, are becoming aware of the benefits of lutein and zeaxanthin for reducing the risk of developing this disease. More recently, scientific studies indicate that lutein and zeaxanthin may also reduce the risk of other major chronic diseases such as breast cancer, skin cancer and cataracts. is the most abundant carotenoid present in green vegetables, such as spinach, kale, broccoli, Brussel sprouts and leaf lettuce. In fruits, lutein are present in small amounts and are esterified with either palmitic acid, estearic acid, oleic acid, mysteric acid or other fatty acids. have not been found in abundance in human serum or plasma (Khachik et al. 1995a, Bone et al., 1993); they must undergo hydrolysis in presence of digestive enzymes before they can be absorbed into the bloodstream (Khachik, et al. 1995b). This technical paper provides actual statistical data demonstrating that most of the lutein absorbed by the human body is present as lutein not lutein in our diets. Methodology and zeaxanthin content in fruits and vegetables /zeaxanthin content in fruits and vegetables was taken from the USDA-NNCC Carotenoid Database for U.S. Foods (1998). This database was created by the United States Department of Agriculture (USDA) and the Nutrition Coordinating Center (NCC) at the University of Minnesota. The project was partially funded by the National Cancer Institute (NCI). Most values for lutein and zeaxanthin were reported as one. Where the zeaxanthin values were available, these values were subtracted from the total lutein/zeaxanthin content and expressed as lutein concentrations. In those cases where zeaxanthin numbers were 80325 Trademarks Registered by Kemin Industries, Inc. Page 1 of 5

not reported, it was assumed that lutein/zeaxanthin values represented lutein. Since lutein is not found in most fruits (Khachik, et al. 1995b), it was also assumed that all the lutein/zeaxanthin values reported in this database for fruits represented lutein and zeaxanthin. Per capita consumption of lutein, zeaxanthin and their in the United States The per capita consumption of lutein, lutein, zeaxanthin and zeaxanthin in the USA for 1996 was calculated based on the per capita consumption of individual fruits, vegetables and eggs. Only fresh and frozen vegetables were included in this research. Cooked vegetables were purposely excluded because the amount of lutein present in these vegetables is dependent on cooking temperatures and rates, which may vary (USDA Statistics reports, 1999) (Figure 1). To estimate the per capita consumption of these carotenoids and their, the per capita consumption for each fruit and vegetable (Figure 1) was multiplied by the average content of lutein, lutein, zeaxanthin or zeaxanthin reported on Table 1. The data are expressed in mg/year. Figure. 1. Per Capita of Fruits, Vegetables and Eggs in the USA (1996). 55 50 (pounds/year) 45 40 35 30 25 20 15 10 5 0 Products Broccoli Brussel Sprouts Cabbage Carrots Celery Corn, sweet Grapefruit, red Green Beans Kale, raw Lettuce, iceberg Leaf Lettuce Melons Oranges Papaya Peaches Peas (Green) Spinach Winter Squash Summer Squash Tangerines Tomatoes Watermelon Eggs From USDA statistics reports (1999) 80325 Trademarks Registered by Kemin Industries, Inc. Page 2 of 5

Results Table 1 shows the distribution of lutein, lutein, zeaxanthin and zeaxanthin in fruits, vegetables and eggs. Most of the lutein from the diet is found in green vegetables, particularly, kale and spinach. According to the data shown on Table 1 most of the per capita lutein intake comes from broccoli, spinach, leafy lettuce and iceberg lettuce. and zeaxanthin are present in very small amounts in fruits. The major contributors of lutein in a typical diet are tomatoes and oranges. This is true because of the high per capita consumption of these fruits per year, however these numbers only represent less than five percent of the total annual per capita consumption of lutein, zeaxanthin and their. Therefore, it is estimated that 80 percent of total per capita consumption of these carotenoids are found in nature as lutein and 13 percent as zeaxanthin (Figure 2). Only small amounts of lutein (5.6 percent) and zeaxanthin (1.4 percent) are present as such in our diets. Table 1. Composition of Fruits, Vegetables & Eggs and Per Capita of,, & (1996). Fresh and frozen vegetables & eggs ester Broccoli 2,422 62.68 - - 23 0.60 - - Brussel Sprouts 1,590 0.79 - - - - - - Cabbage 310 14.36 - - - - - - Carrots, baby 335 21.29 - - 23 1.46 - - Celery 229 6.71 - - 3 0.09 - - Corn, sweet 356 19.72 - - 528 29.24 - - Grapefruit, red - - 13 0.34 - - - - Green Beans 596 7.04 - - 44 0.52 - - Kale, raw 39,377 1.79 - - 173 0.01 - - Lettuce, iceberg 282 31.11 - - 70 7.72 - - Leaf Lettuce 2,448 67.79 - - 187 5.18 - - Melons - - 40 0.42 - - - - Oranges - - 113 6.57 - - 74 4.30 Papaya - - 75 0.14 - - - - Peaches - - 51 1.20 - - 6 0.14 Peas (Green) 1,292 8.21 - - 58 0.37 - - Spinach 11,607 68.50 - - 331 1.95 - - Winter Squash - - 38 0.01 - - - - Summer Squash 2,125 1.54 - - - - - - Tangerines - - 131 1.13 - - 112 0.97 Tomatoes - - 130 11.15 - - - - Watermelon - - 17 1.34 - - - - Eggs 32 7.55 - - 23 5.43 - - - - - - - - - - Total 63,401 319 608 22 1,463 53 192 5.4 From USDA-NNCC Carotenoid Database for U.S. Foods (1998) * These numbers were calculated based on the per capita consumption of and the amount of the carotenoids present in the products. 80325 Trademarks Registered by Kemin Industries, Inc. Page 3 of 5

Figure 2. Prevalence of and in Fruits, Vegetables and Eggs in Human Diet. Nebeling, et al (1997). 13% 6% 1% 80% Nebeling, et al (1997) estimated that the average intake of /zeaxanthin was 2 mg/day. These calculations also included the contribution of lutein and zeaxanthin. However, based on the distribution of these carotenoids and their in fruit, vegetables and eggs, these only contribute to seven percent of the total intake of lutein and zeaxanthin, meaning that on average, people only consume 0.14 mg/day of lutein and zeaxanthin in their diet (Figure 2). Furthermore, because lutein and zeaxanthin must be de-esterified by the body in order to be absorbed, it is not clear the actual contribution to the total lutein content found in serum and deposited in body tissues these make. It is also worthwhile mentioning that the results from this data are consistent with the low daily average consumption of lutein and zeaxanthin previously reported by Nebeling, et al. (1997). Furthermore, these numbers are much lower than the suggested lutein intake of 6 mg/day reported in the Journal of American Medical Association in 1994 by Dr. Seddon and co-workers. These researchers found a high correlation of AMD-disease prevention with an intake of 6 mg/day of lutein from spinach and other dark leafy vegetables. Therefore, they recommended a diet rich in lutein to lower the risk of developing AMD (Seddon, et al. 1994). 80325 Trademarks Registered by Kemin Industries, Inc. Page 4 of 5

Conclusions The results of this research demonstrate that 93 percent of the total amount of lutein, zeaxanthin and their found in fruit, vegetables and eggs are present as lutein and zeaxanthin and not as lutein or zeaxanthin. This is an important consideration when choosing to supplement our diets with lutein and zeaxanthin because some ingredients currently available in the market contain lutein and zeaxanthin. As this research demonstrates, lutein and zeaxanthin, as found in marigold extract, are not the predominant sources of lutein and zeaxanthin found in a typical diet. References 1. Bone, R. A., J. T. Landrum, et al. (1993). "Stereochemistry of the human macular carotenoids." Invest Ophthalmol Vis Sci 34(6): 2033-40. 2. Khachik, F., G. R. Beecher, et al. (1995). ", lycopene, and their oxidative metabolites in chemoprevention of cancer." J Cell Biochem Suppl 22: 236-46. 3. Khachik, F., N. Zohar, et al. (1995). Distribution of carotenoids in fruits and vegetables as criterion for the selection of appropriate chemopreventive agents. International Conference on Food Factors - Chemistry & Cancer Prevention, Hamamatsu, Japan. 4. Nebeling, L. C., M. R. Forman, et al. (1997). "Changes in carotenoid intake in the United States: the 1987 and 1992 National Health Interview Surveys." J Am Diet Assoc 97(9): 991-6. 5. Seddon, J. M., U. A. Ajani, et al. (1994). "Dietary carotenoids, vitamins A, C, and E, and advanced age-related macular degeneration. Eye Disease Case-Control Study Group." JAMA 272(18): 1413-20. 6. USDA Statistics Reports (1999). From the United State Department of Agriculture Annual Statistical Reports www.usda.gov 7. USDA-NNCC Carotenoid Database for U.S. Foods (1998). www.usda.gov 80325 Trademarks Registered by Kemin Industries, Inc. Page 5 of 5