Download Glucoraphanin - An Indirect Antioxidant Found Naturally In Broccoli

G E N E R A L I N F O R M AT I O N
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Glucoraphanin - An Indirect Antioxidant Found
Naturally In Broccoli That Supports Cell Integrity
And Protects Against Free Radical Damage
By Jeremy Bartos, Ph.D.
Scientific & Regulatory Affairs Manager
Glanbia Nutritionals
Glanbia Nutritionals | truebroc® White Paper | June 2015
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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introduction
Today’s health-conscious consumers are more informed and want to
understand what they are eating as well as the benefits of the products
they consume. They seek natural ingredients that have no additives or
preservatives and are botanical or organic in nature. Currently, there are a
large volume of products on the market that have an antioxidant health
claim. In 2015 alone almost 1000 new products with an antioxidant
position have been released1, but how well do they really work? Recent
scientific research on broccoli has revealed a “next generation” antioxidant
that will potentially change the way we choose and utilize our ingested
antioxidants. truebrocTM glucoraphanin, a naturally derived phytonutrient
extracted from broccoli seeds, is a rechargeable antioxidant that works
with the body’s own cellular protection system. Currently, the most widely
accepted source of ingested antioxidants are short-term “one and done”
antioxidants, such as Vitamins C and E and polyphenols such as
resveratrol (grapes and wine) and EGCG (green tea) that last for only a
short time in the body. The effects of truebrocTM have the benefit of lasting
several days in the body – long after the “one and done” antioxidants have
dissipated. truebrocTM glucoraphanin, the next generation of antioxidants,
activates a sustainable antioxidant system within the body that is
rechargeable and meets the demands of today’s health conscious
consumers who are looking for “clean label” options with clearly stated
benefits.
PRODUCTS WITH ANTIOXIDANT
CLAIMS ON THE RISE
WH AT M A KES TRUE BROC T M A PRE MIE R
ANTIOXIDANT
> truebrocTM boosts Phase II Enzymes, enhancing the
body’s own removal of free radicals and overall
detoxification of cellsNon-burning and non-irritating
to the stomach
> Standardized to 13% glucoraphanin – highest
concentration available
> Made from selected natural broccoli seeds grown
in California and water extracted in Canada
• Complete traceability from field to product
• Produced under cGMP
> Patent Protected – source and process
> Self-Affirmed GRAS
> Recommended dosage:
• Supplements – 30mg of glucoraphanin
(230mg of truebrocTM)
• Foods – 10-15mg of glucoraphanin (77115mg of truebrocTM)
> Applications:
• Tablets
• Capsules
• Powders
• Functional Beverages
• Functional Foods
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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THE IMPORTANCE OF ANTIOXIDANTS
Everyone has heard about antioxidants and knows that they are good for them, but does everyone really know how antioxidants work in
our bodies? The main purpose of antioxidants is to protect the cells in the body from damage caused by oxidative stress, namely free
radicals, which are unstable compounds missing an electron (a negative charge) on their outside layer2. Free radicals are both created as
a byproduct of normal cellular respiration as well as environmental byproducts of pollution, sunlight, etc. Because of their instability they
“steal” electrons from any source they can find in the body to help stabilize them. The most readily available source of electrons in the
body is the phospholipid bilayer that surrounds each cell. Over time, the chronic exposure of the cell walls to free radicals will break
down the walls, which can have a variety of negative effects, both acute and chronic. In some cases it is a major cause of a disease,
such as seen in Alzheimer’s3. In others, it simply makes an already existing condition worse, as in asthma or Rheumatoid Arthritis4,5,6,7.
By definition, antioxidants are electron donors; they act as sacrificial lambs by offering free radicals another source of electrons besides
the cell wall. They can give their electrons to free radicals, neutralizing them and thereby preventing them from stealing any more
electrons and doing further damage. The antioxidants can thus help prevent or minimize the build-up of damage over time.
Unfortunately, free radical damage is also closely linked with inflammation because the body recognizes the oxidative damage as a
threat8. To answer this, the body sends out an army of soldiers in the form of immune cells. These immune “soldiers” fight against
anything they perceive to be foreign and once they are done, like casualties in any battlefield, there will be a mass of dead cells. This in
itself sends signals for an influx of more immune cell soldiers. What results is a cycle of oxidation - damage – immune cells - more
damage. Increasing antioxidant potential in the body provides a solution by helping to prevent a lot of the oxidative damage in the first
place and thus also helps minimize localized inflammation, for example in the muscles after rigorous exercise9, 10. Without antioxidants,
our cells could be easily damaged by reactive oxygen species, DNA damaging electrophiles, inflammation and radiation.
The body has several ways of removing free radicals before they can cause damage. The method best known to consumers is to ingest
antioxidants through food or supplements, such as Vitamin C and polyphenols11,12. Slightly less known are native antioxidants and
enzymes produced by the body, such as glutathione and superoxide dismutase13,14,15. And finally there is the Phase II Enzyme System;
enzymes that act to increase the body’s native antioxidant and detoxifying pathways. The Phase II Enzyme System is activated by Nrf2,
a transcription factor that, when induced, increases the production of the specific antioxidant enzymes16,17,18.
LONG-LASTING ANTIOXIDANT EFFECTS
Most dietary antioxidants, such as vitamins and polyphenols,
act by donating electrons to free radicals. Once they do this,
their antioxidant function is depleted: they work in a “one-anddone” fashion. The production of free radicals continues and if
the supply of one-and-done antioxidants runs out then those
free radicals will start stealing again, which can lead to oxidative
damage. Figure 1 demonstrates the “one and done” effect of
various amounts of ingested Vitamin C; unless the body gets
constant replenishment (as often as 4 to 6 times a day) of these
ingested antioxidants, the antioxidant protection quickly wanes
in a matter of hours19.
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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Native antioxidants such as glutathione and superoxide dismutase are “rechargeable” antioxidants; once they have donated electrons
they can be “recharged” by the members of Phase II Enzyme system, thus they can be reused over and over again20,21. One way to
ensure this occurs is by activating the Phase II Enzyme system; without it, native antioxidants are also just “one and done”. The major
regulator of the genes that code the Phase II Enzymes is nuclear factor erythroid 2-related factor 2 (Nrf2), which binds the antioxidant
response element (ARE) region of the gene promoter. This in turn leads to the expression of antioxidant and Phase II Enzymes,
demonstrating that Nrf2 is the key to start this Phase II Enzyme system22. Therefore, compounds that can activate Nrf2 can indirectly
activate the entire Phase II Enzyme system23. Once activated, the Phase II Enzymes can remain operational for as long as 72 hours,
thereby ensuring long-lasting systemic antioxidant effects24. In addition to Phase II Enzyme regulation, Nrf2-ARE binding regulates the
expression of more than 100 other genes involved in cellular antioxidant and anti-inflammatory defense such as heat shock proteins and
ferritin, pro- and anti-inflammatory enzymes such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and heme
oxygenase-1 (HO-1)25,26. It has been shown that down- regulation of Nrf2 can have a variety of negative effects, from increased oxidative
stress load and environmental toxin levels to localized inflammation and aberrant mitochondrial biogenesis27,28. Keeping this important
pathway up and running is critical to maintaining healthy cellular processes and staving off chronic maladies.
DISCOVERY: A NEW ANTIOXIDANT COMPOUND FOUND IN BROCCOLI
In the early 1990’s, scientists at Johns Hopkins University identified a compound derived from broccoli called sulforaphane29,30.
Sulforaphane and its precursor glucoraphanin (sulforaphane glucosinolate) act as part of the plant’s defense system; in fact,
sulforaphane is responsible for the characteristic sulfur smell/taste of broccoli. Glucoraphanin belongs to a category of compounds
called glucosinolates, which are naturally found in cruciferous vegetables. It is converted into the isothiocyanate sulforaphane by an
enzyme found in broccoli called myrosinase or in the body’s gut microflora31. In the broccoli plant, myrosinase and glucoraphanin are
kept in different cellular compartments. When the plant is chewed or cut the myrosinase and glucoraphanin can interact, resulting in the
conversion of glucoraphanin into sulforaphane. It is the sulforaphane that has the characteristic bitter broccoli taste designed to
discourage animals from eating the plant! In the course of their experiments, the JHU scientists discovered that sulforaphane was one of
the most potent activators of Nrf2 found in nature29.
This led to a flurry of publications demonstrating the effect that
sulforaphane, glucoraphanin, and broccoli extracts have on the
activation and expression of specific Phase II Enzymes32. Figure 2
demonstrates the effect of sulforaphane on the activation of several
of these enzymes24. Quinone Reductase (QR) helps to reduce
electrophilic quinones, an important step in cellular detoxification.
Glutathione Reductase (GR) catalyzes the reduction of glutathione
disulfide (uncharged) to glutathione (charged). Glucose-6Phosphate Dehydrogenase (G6PD) helps maintain the cellular level
of NADPH which in turn maintains the levels of glutathione.
Glutathione (GSH) itself is an indirect target that also increases due
to increased activity of Glutathione Reductase.
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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SULFORAPHANE CLINICAL STUDIES SHOW
POSITIVE RESULTS
Recent human studies on sulforaphane and broccoli products have also
shown positive results, similar to those seen in the animal trials. A 17
subject randomized, crossover dietary trial was set up to examine the
effect of standard broccoli (S) and high-glucoraphanin (HG) broccoli
(broccoli with three times more glucoraphanin than standard broccoli)
as a single meal on the expression of key Phase II Enzymes, compared
with water (w)33. Thioredoxin reductase 1 (Tr1) is an enzyme critical to
the thioredoxin pathway that “recharges” thioredoxin, an antioxidant
similar to glutathione that acts as an electron donor to peroxidases and
ribonucleotide reductase to help reduce free radical damage.
Glutamate-cysteine ligase modifier subunit (GCLM) is a subunit of
gamma-glutamylcysteine synthetase, the first rate-limiting enzyme of
glutathione synthesis. As seen in Figure 3, high-glucoraphanin broccoli
showed a >2x induction of these key antioxidant Phase II Enzymes over
standard broccoli controls and a >20 fold induction over water33.
In another human crossover study, 20 subjects were given a 4 week
intervention of either 2 cups broccoli (~218 grams; BV) or a supplement
containing fiber and antioxidant micronutrients found in broccoli such as
60 mg Vitamin C and 5000 IU Vitamin A (M+F)34. In this study, they
measured changes in F2-isoprostanes (F2iP), which are biomarkers that
are becoming widely recognized as a highly reliable index of body-wide
oxidative stress and are easily measurable in urine samples. It was
found that consuming 218 grams of broccoli/day decreased overall
oxidative stress markers by an average of 22%, while the antioxidant
vitamins showed no change in isoprostane levels, demonstrating that
the glucoraphanin is the key driver for the effects seen (Figure 4).
In a human dose-escalation placebo-controlled trial, 65 subjects were spread out
over 5 groups and given different amounts of broccoli sprout homogenate; a
placebo, 50 g broccoli sprout homogenate (BSH), 100 g BSH, 150 g BSH, 200 g
BSH35. The aim of this study was to determine if the sulforaphane from broccoli
stimulates the production and activation of Phase II Enzymes in a dose dependent
manner. In this case, they measured changes from baseline in the expression of
NADPH quinone oxidoreductase (NOQ1), which reduces hydrogen peroxide free
radicals (a toxic byproduct of cellular respiration), and glutathione S transferase P1
(GSTP1), which functions in removal of free radicals and acts in concert with
tumor suppressing protein p53. As highlighted in the circles in Figure 5, it was
shown that sulforaphane does indeed increase expression of these Phase II
Enzymes in a dose dependent manner, as people who consumed lower amounts
of BSH per serving showed lower expression than those people who consumed
higher amounts of BSH per serving. Therefore, the more sulforaphane you take,
the better the response!
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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THE ALTERNATIVE TO EATING LARGE AMOUNTS OF BROCCOLI
Who wants to eat 200 grams of broccoli sprout homogenate? Just because broccoli is
good for you doesn’t mean people want to eat it, especially at levels high enough to have
a meaningful effect on their health. In fact, in an online omnibus survey of 1,012 nationally
representative U.S. adults ages 18+ conducted in March 2015, nearly 1 in 3 (32%)
Americans have found a reason to avoid eating more broccoli36. Furthermore, nearly 2 in
5 (39%) wish they could get the nutritional benefits from broccoli without actually having
to eat it! Therefore, this shows that there is a large group of people who are looking for an
easier way to get the antioxidant boosting benefits of broccoli.
WHY TRUEBROCTM IS THE ANSWER
Enter truebrocTM, a patented and highly concentrated broccoli
seed extract standardized to glucoraphanin, born from the same
Johns Hopkins University scientists who first identified the
antioxidant benefits of sulforaphane from broccoli. And this is
important because broccoli is variable. The levels of the actives
in broccoli vary wildly due to genetic variations, the location
where it’s grown, and on the part of the plant consumed (florets,
stalks, sprouts, seeds, etc.)37,38. In addition, the broccoli florets
found in your grocery store have less glucoraphanin than broccoli
sprouts and seeds. It has been determined that broccoli seeds
have significantly higher levels of glucoraphanin than both
broccoli florets and broccoli sprouts, which further translates into
higher Phase II Enzyme Inducer Activity (Figure 6, data not
published). The proprietary truebrocTM seeds are grown in
California and the glucoraphanin is water-extracted under
patented cGMP processes in Canada with complete traceability
field to product. Almost twenty years of research into harvesting and extraction has resulted in truebrocTM, standardized to 13%
glucoraphanin, the highest level on the market. A 230 mg daily serving of truebrocTM contains as much glucoraphanin as 10 ounces of
regular broccoli – more than half a pound!
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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AS NATURE INTENDED IT
Plants do not naturally produce sulforaphane; they make the components required to synthesize sulforaphane when the conditions are
right (glucoraphanin and myrosinase). This is because sulforaphane is known to be highly unstable, especially in high temperature
conditions and as such would degrade over time if the plant made and stored it.39,40 Therefore one of the main reasons to standardize to
glucoraphanin versus converting it to sulforaphane is because of the stability of glucoraphanin, especially in high heat conditions, with
only ~ 5% losses as a result of cooking or steaming41. Sulforaphane stability issues can be further exacerbated in finished products such
as supplements and skin care products, where temperature sensitivity can translate into reduced shelf-life, especially in hotter
environments. It was shown that the degradation rate of sulforaphane increased by a factor of nearly 3.1 for every 10 °C increase in
temperature with complete degradation seen after 30 days in a conventional pharmaceutical cream formulation42. Advanced spray
drying techniques may help stabilize sulforaphane for extended periods of time but will add cost while ultimately diluting the product43. In
addition, the bitter flavor profile of sulforaphane is also not easy to mask, making glucoraphanin a more pleasant candidate for use in
foods, beverages, and the like.
TRUEBROCTM BIOEQUIVALENCY STUDIES PROVE IT’S EFFECTIVE
The patented harvest and extraction processes used for truebrocTM builds on the original work performed at Johns Hopkins University.
The majority of the original JHU research utilized broccoli sprouts standardized to the levels of glucoraphanin compared to broccoli
florets. Further analyses led to the discovery that broccoli seeds were even higher in glucoraphanin than sprouts, providing the
foundation for the seed cultivation that would lead to the unique proprietary broccoli seeds extracted to make truebrocTM. A crossover
study was performed at Johns Hopkins University to determine if the conversion efficiency of glucoraphanin to sulforaphane is the same
when truebrocTM is used compared to the 3-day old broccoli sprout preparations that had been used in many JHU studies44. The urine
samples of 20 adults were analyzed for sulforaphane metabolites 4 separate times using two different doses of two different
glucoraphanin preparations; two from truebrocTM and two from 3-day old broccoli sprouts prepared in the same manner as used in all of
the JHU studies. When the values for the 20 subjects were averaged, the percent conversions of glucoraphanin to sulforaphane for the
two preparations (truebrocTM and broccoli sprouts) were statistically the same and in the range of 11% to 13% for the lower dose and
8% to 10% for the higher dose. This study demonstrated the bioavailability of glucoraphanin from truebrocTM is equivalent to that of the
3-day-old broccoli sprout preparations that have been used in all Johns Hopkins University studies, thus providing evidence that all of
the results seen in the Hopkins studies would be similar when using truebrocTM.
CONCLUSION: TRUEBROC TM IS A PREMIER LONG-LASTING INDIRECT ANTIOXIDANT THAT
SUPPORTS CELL INTEGRITY AND PROTECTS AGAINST FREE RADICAL DAMAGE
truebrocTM has many advantages over more widely consumed and recognizable antioxidants because the glucoraphanin in truebrocTM is
converted into sulforaphane by gut microflora and, once converted, it boosts the body’s own Phase II Enzyme System to help protect
cells. This system helps to reduce oxidative stressors and enhance the removal of environmental toxins, increasing cellular antioxidant
defense and contributing to healthy organ function.
The addition of truebrocTM benefits brand owners and manufacturers seeking a “true” antioxidant with beauty-from- within, sports
nutrition, cellular protection and overall heightened antioxidant capabilities.
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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WHY GLANBIA NUTRITIONALS?
Glanbia Nutritionals is a global leader in ingredient solutions,
providing precision premixes, amino acids, vitamins, minerals,
specialty ingredients and colors for the food, beverage and
supplement industries. Built on our reputation for outstanding
quality and service, we deliver formulation and ingredient
expertise that help our customers optimize their products and
propel them to greater success.
Glanbia Nutritionals
2840 Loker Avenue East
Carlsbad, CA 92010
+1 800 735 8137
[email protected]
glanbianutritionals.com
© 2015 Glanbia Nutritionals. All rights reserved.
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37. Saha et al; Mol Nutr Food Res. 2012 Dec;56(12):1906-16;
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44. Paul Talalay, P.I., Conversion of Broccoli Seed Glucosinolates to Isothiocyanates – Dietary Supplements, “submitted 08/28/2014
Glanbia Nutritionals | truebroc® White Paper | June 2015
These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure or prevent disease.
Glanbia Nutritionals | 5951 Mckee rd., Suite 201 | Fitchburg, WI 53719 | 800.336.2183 | 608.316.8500 | www.glanbianutritionals.com
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