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Tracy Beckmann Dietary Analysis April 18, 2011 Dietary Analysis for the Broccoli Sprout Study 2010 The goal of this study was to examine the absorption, metabolism and activity of sulforaphane derived from either broccoli sprouts or a purified broccoli extract supplement in human volunteers. The cross-sectional study recruited healthy adult subjects (n=16) aged 18-60 years old. For the first phase of this study (completed in early April 2010), subjects consumed 40 g of broccoli sprouts (alfalfa sprouts for control group) then provided blood and urine samples at 0, 3, 6, 12, 24, and 48 hours post-broccoli sprout consumption. Subjects kept dietary records for three days (each phase) to monitor dietary intake. For one week prior to and during each phase of the study, subjects refrained from eating vegetables containing organosulfur compounds (including but not limited to broccoli, cabbage, onions, garlic and kale). To compare the differences in metabolism of sulforaphane from whole food versus supplement intake after a one-month washout period, in the same subjects, the above procedures were repeated with a broccoli extract supplement (or placebo). For both phases of the study metabolites in blood and urine samples and epigenetic markers in blood samples were analyzed. This study proposes to increase knowledge of sulforaphane metabolism as well as to provide evidence supporting dietary broccoli consumption, in whole food versus supplement form, as an effective means of reaching physiological levels of sulforaphane which may exert anti-cancer effects. Dietary intake data compiled from the Broccoli Sprout Study 2010 combines information provided from subject food records over 2 phases, each 3 days long, for 6 days total. Multivitamin and mineral intakes were included in the nutrient analyses. Food record information was entered into The Food Processor SQL Dietary Analysis Software, 2010, Version10.6.0, available from ESHA Research, Salem, OR. Analysis of both subject macronutrient and micronutrient intake was evaluated (see Table 1 for specific nutrients analyzed). Between Phase 1 and 2 of the study, the difference in the average intake of all nutrients was ≤ 12.5%. Therefore, subject dietary intake was consistent between phase 1 and 2 of the study. Table 1: Nutrient Intake for Broccoli Sprout Study 2010. Nutrient intakes of subjects compared to the Recommended Dietary Allowances (RDAs) for all macronutrients and several micronutrients. Nutrient Calories (kcal) Protein (g) Carbohydrates (g) Fat (g) Phase 1 Mean Intake 2214 86 302 76 Phase 1 Percent of RDA 84% 141% 83% 92% Phase 2 Mean Intake 2304 90 321 72 Phase 2 Percent of RDA 88% 148% 89% 89% Vitamins Vitamin A (RAE) Vitamin B1 (mg) Vitamin B2 (mg) Vitamin B3 (mg) Vitamin B6 (mg) Vitamin B12 (mcg) Vitamin C (mg) Vitamin D (IU) Vitamin E (mg) Folate, DFE (mcg) 864 2 2 25 2 5 168 384 13 725 114% 180% 193% 165% 157% 210% 205% 64% 87% 173% 774 2 2 25 2 6 147 420 13 639 102% 169% 194% 170% 170% 264% 183% 70% 87% 152% Minerals Calcium (mg) Iron (mg) Potassium (mg) Sodium (mg) Average caloric intake consumed during Phase 1 and Phase 2 of the study was 84% and 88%, respectively, of the Recommended Dietary Allowances (RDAs) as calculated by The Food Processor software, which takes into account age, gender and activity level. A similar pattern was observed for fat and carbohydrate intake. Fat intake was 92% for Phase 1 and 89% for Phase 2 of the RDA. Carbohydrate intake was 83% for Phase 1 and 89% for Phase 2 of the RDA. Accuracy of food records is affected by the subject’s aptitude to accurately record the portion size consumed and the tendency to underreport the actual amount of food consumed (1). An individual may also alter the amount of food consumed as a result of the food recording process (1). In addition to the inaccuracy of food records, further error is introduced into the dietary analysis by limitations of the nutrient analysis programs/databases. Some food products are not found in the database; therefore, closely matched, but not exact, products are used in analysis. Also, many products in the database have limited or incomplete data, especially for micronutrients. Despite the above mentioned limitations, food records are a cost effective method and provide detailed daily food and nutrient intake data, including food preparation method and snack and meal timing (1). 1040 24 2183 3321 103% 198% 46% 169% 1178 26 2390 3164 117% 218% 51% 159% Several other factors may also have affected food record data. One subject was on a weight loss diet and consumed approximately 61% of recommended caloric intake. However, the same diet plan was followed for both phases of the study. Two subjects had highly variable food consumption with caloric intake differing by 700-1300 kcals from one day to the next. This variability may indicate inconsistent recording. In addition, there was incomplete vitamin and mineral supplementation information for several of the subjects in this study. The study criteria included restriction of cruciferous vegetables for one week prior to and during each phase of the study. One subject did not refrain from cruciferous vegetable intake for the week prior to Phase 1 and a second subject consumed an order of onion rings during Phase 2 Day 1 dinner. Average protein intake was higher than the RDA with amounts of 141% for Phase 1 and 148% for Phase 2. These results are consistent with current literature on the average amount of protein consumed by Americans, which states intake at 86 g, or 141% (2). Subject intake of protein between the two phases of this study maintained <5% difference. Most micronutrients were consumed at the recommended amounts or higher except for vitamins E, D and potassium. For micronutrients that were consumed in amounts exceeding 100% of the RDA (including vitamins A, B1, B2, B3, B6, B12, C, folate, calcium, iron and sodium), the levels consumed were well below the Upper Limit (UL) for those nutrients. The average intake of vitamin E for both phases was 13 mg, or 87% the RDA for potassium. In the US, average vitamin E intake is 50% or less (8 mg/d for men, 6 mg/d for women) of the RDA (15 mg) (3). Subjects in our study consumed more vitamin E than the average American. Overall, reported vitamin E consumption may be underestimated due in part to unknown vitamin E levels of cooking and other oils consumed (4). The mean vitamin D intake from foods alone among subjects was 64% of RDA for Phase 1 and 70% of RDA for Phase 2. The recommended amount for vitamin D was increased in 2010 to 600 IU per day for all adults aged 18-60 years old (5). The National Institutes of Health estimate the average intake of vitamin D by Americans to be 34%-48% of RDA per day for males and 24%-46% of RDA per day for females (6). Subjects in this study consumed more vitamin D than the average American. Although vitamin D is found naturally in relatively few foods, it is widely available in fortified foods (especially dairy) and in supplement form (6). Additionally, vitamin D is converted to its active form in the human body via exposure to ultraviolet light (5). Potassium intake was at 46% and 51% of recommended for Phase 1 and 2, respectively. Western diets are typically lower in potassium than the RDA, thought to be due to the high sodium content of such diets (the processing of foods generally increases sodium levels and decreases potassium levels) (8). Inadequate potassium intake is relatively common as the RDA is set at a level for prevention of chronic disease, so even though the RDA is not being met deficiency is unlikely. The modern Western diet, which consists of many processed foods and lacking in whole grains, fresh fruits and vegetables, is therefore high in sodium and low in potassium and may be a factor in several chronic diseases (7). Analysis of dietary intake for the Broccoli Sprout Study 2010 found that subject intake of nutrients was consistent and caloric contents similar during both phases of the study. All subjects met their RDAs for age and gender with the exception of vitamins E, D and potassium, which were consumed in lower amounts. Minimal to no sulforaphane foods were consumed during the two study periods. Analysis of subject food records indicates that subjects were nourished and adhered to the dietary recommendations of the study, and may offer insight to any confounding results of sulforaphane absorption, metabolism and activity. References: 1. Mahan, L. K., and Escott-Stump, S. (2008.) Chapter 14: Assessment: Dietary and Clinical Data. Krause’s Food and Nutrition Therapy, 12th Edition, pp. 395-399. Missouri: Saunders, Elsevier. 2. National Center for Health Statistics (NCHS). (2010.) Adult’s Protein Intake Much More Than Recommended. Retrieved from http://nchspressroom.wordpress.com/category/nhanes/ on 3/23/11. 3. Linus Pauling Institute. (2007). Oregon State University. Micronutrient Information Center: Vitamin E. Retrieved from http://lpi.oregonstate.edu/infocenter/vitamins/vitaminE/ on 3/26/11. 4. Office of Dietary Supplements.(n.d.). National Institutes of Health. Dietary Supplement Fact Sheet: Vitamin E. Retrieved from http://ods.od.nih.gov/factsheets/vitamine/. Accessed on 11/15/10. 5. Linus Pauling Institute. (2010). Oregon State University. Micronutrient Information Center: Vitamin D. Retrieved from http://lpi.oregonstate.edu/infocenter/vitamins/vitaminD/index.html on 3/26/11. 6. Office of Dietary Supplements.(n.d.). National Institutes of Health. Dietary Supplement Fact Sheet: Vitamin D. Retrieved from http://ods.od.nih.gov/factsheets/vitamind/ . Accessed on 3/25/11. 7. Linus Pauling Institute. (201). Oregon State University. Micronutrient Information Center: Potassium. Retrieved from http://lpi.oregonstate.edu/infocenter/minerals/potassium/ on 3/26/11. 8. Thompson, J.L., Manore, M. M., and Vaughan, L.A. (2008.) Chapter 9: Nutrients Involved in Fluid and Electrolyte Balance. The Science of Nutrition, 1st Edition (pp 370-371). San Francisco, CA: Pearson/Benjamin Cummings.