Broccoli Sulforaphane Bioavailability With Mustard
Condition: Sulforaphane Bioavailability
Study type: Interventional

Summary: 
      We know that broccoli is a vegetable that is associated with a number of health benefits and
      is a good choice in a healthy diet. Researchers at major universities, including Johns
      Hopkins University, have identified nutrients in broccoli that have specific health benefits
      such as improving antioxidant protection. One of these nutrients is called glucoraphanin
      (GR). However, in order for GR to deliver health benefits in the human body, it first has to
      be converted to its active form, sulforaphane (SF). This conversion can happen one of two
      ways. GR can be converted to SF by our gut bacteria (the microbiota) or it can be converted
      to SF by an enzyme called myrosinase that is naturally occurring in certain plants, including
      mustard (the same mustard that is used to make table mustard spread and mayonnaise). The
      purpose of the present study is to test in healthy human volunteers the oral bioavailability
      (absorption) of GR in broccoli seed extract (BSE) with or without the presence of active
      plant myrosinase in mustard seed powder (MSP) delivered in a nutritional supplement.
    

Detailed description: 
      The aim of the present study is to determine in healthy human volunteers the oral
      bioavailability of sulforaphane (SF) derived from broccoli seed extract (BSE) standardized to
      glucoraphanin (GR) content with and without the presence of active plant myrosinase derived
      from mustard seed powder. Once absorbed, SF is rapidly metabolized by initial conjugation
      with the endogenous nucleophile glutathione (GSH). The glutathione conjugate then undergoes
      successive steps of hydrolysis leading ultimately to the formation of the N-acetylcysteine
      derivatives (mercapturic acids). All of these conjugates are chemically designated
      dithiocarbamates (DTC) and can be quantified in the blood and urine by mass spectroscopy
      (MS). Based on previous published studies, it is expected that the conversion of the oral
      dose of 115 umol of GR to DTC will be about 30-40% (on a molar basis) when GR is co-delivered
      with mustard seed powder myrosinase, but only 10% when delivered without the myrosinase
      source. Blood samples will be collected to determine if the supplements influence biomarkers
      for oxidative stress and inflammation. The stool sample will be used to measure the
      alpha-diversity of the gut microbiome and whether this has an influence on urine levels of SF
      and DTC.

      This study will include 16 male and female adults ages 25 to 65 years, and employ a
      randomized crossover design, with a 7 day washout period between trials. Supplements
      (broccoli seed extract, BSE, and vitamin C (VC) with or without mustard seed powder, MSP)
      will be in identical capsules, with the double blind code held by the sponsor until after the
      study is over.

      Participants will be recruited and then contacted prior to visit #1 to determine if they
      match the criteria for entry into the study. Participants will record food and beverages
      consumed during the evening meal prior to visit #1. Participants will also be instructed to
      refrain from consuming cruciferous vegetables and condiments that might contain
      glucosinolates or isothiocyanates (e.g., mustard, horseradish, wasabi, mayonnaise) for 3 days
      before and during the duration of the study.

      Visit #1: Screening, Orientation, Supplementation and Basic Study Procedures Participants
      will come to the Human Performance Lab (600 Laureate Way, North Carolina Research Campus,
      Kannapolis, NC) at 7:00, 8:00, or 9:00 am after an overnight fast (at least 9 hours) having
      avoided consumption of cruciferous vegetables and condiments for at least three days. This
      visit will begin with a review of the consent form. Height, body weight, body composition
      (i.e., percent body fat measured on a special scale that participants stand on with bare
      feet), and blood pressure (automated unit, Omron Healthcare, Inc. Lake Forest, IL,
      IntelliSense Blood Pressure Monitor Model: HEM-907XL) will be measured in a private section
      of the lab. The food record for the prior evening meal will be turned in and reviewed with
      instructions to consume the same meal the evening before Visit #3. Participants will next
      provide a spot urine sample in a private stall in the restroom. A mid-stream urine sample
      will be collected in a cup. Next, participants will be randomized to consume 2 capsules
      containing either BSE + VC or BSE+VC+MSP along with 8 oz. of water. Participants will consume
      all of the water along with the 2 capsules. The two supplements will be in identical looking
      capsules so that participants will not be able to determine their identity. Participants and
      the research will not know which capsules were received at visits 1 or 3 until the study is
      over. Following intake of the supplement, participants will consume a muffin and coffee
      within 15 minutes of taking the supplement. Participants will abstain from eating any food or
      beverages except for water for 2 hours following consumption of the supplement. After the 2-h
      delay, participants can resume their normal dietary patterns. Participants will collect all
      urine for the 24 h period after consuming the supplements. Participants will collect all
      urine for the first 8 hours in one container, and then collect the remaining 8-24 h of urine
      in a second container. This first visit will take about 1 hour.

      Visit #2: Return urine samples and provide blood sample Participants will come to the lab in
      an overnight fasted state the next morning, drop off the urine collection containers, and
      provide a blood sample. A stool collection kit with instructions will be given to each
      participant. (Instructions are included in the forms file attached to this application).

      One Day Prior to Visit #3 Participants will collect a small swab sample of stool in a
      container provided by the lab on Visit #1. Participants will consume the same dinner that was
      consumed prior to Visit #1.

      Visit #3: Supplementation and Basic Study Procedures One week after visit #1, participants
      will return to the Human Performance Lab at the same time in an overnight fasted state. All
      the procedures from Visit #1 will be repeated except that participants will consume the
      opposite supplement.

      Visit #4: Return urine samples and provide blood sample Participants (overnight fasted state)
      will drop off the final urine collection containers at the lab the next morning and provide
      the fourth and final blood sample.
    

Intervention name(s): Broccoli seed extract; Broccoli seed extract with mustard seed powder
Intervention type(s): Dietary Supplement; Dietary Supplement
Intervention description(s): 2 capsules, one dose; 2 capsules, one dose
Intervention MeSH terms: Mechlorethamine
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: Randomized crossover with a 7 day washout period between trials.
Masking: Double (Participant, Investigator)
Primary purpose: Basic Science
Keywords: broccoli; mustard; sulforaphane; glucoraphanin; bioavailability