Here are the results of safety testing conducted at the Knoebel Institute for Healthy Aging (KIHA), University of Denver and  Steep Hill Laboratories in Honolulu, Hawaii. We know Greska’s Carbon-60™ was safe for human consumption, as we are 100% certain our proprietary non-solvent method of producing our C60 uses NO SOLVENTS at any step of the manufacturing process. We then add our C60 to high quality food grade organic sunflower oil. In order to provide the best quality product to our customers, our objective was to verify and provide documentation to our customers that Greska’s Carbon-60™ was not toxic. We wanted confirmation, that not only was Greska’s Carbon-60™ manufacturing process non-toxic but there were no other toxic substances in our product.

PART ONE FOR TESTING GRESKA’S CARBON-60™ FOR TOXICITY

Part One of testing of Greska’s Carbon-60™ for toxicity was conducted at the Knoebel Institute for Healthy Aging (KIHA), University of Denver. Located at the base of the Rocky Mountains in Denver, Colorado, KIHA was founded in 2010 when DU received a generous gift from Betty Knoebel. Mrs. Knoebel was the widow of Ferdinand “Fritz” Knoebel, who was a pioneer in the food services industry. KIHA’s mission is to create and implement solutions for aging issues through multidisciplinary research, education and outreach. Greska’s Carbon-60™ testing was led by Daniel Paredes, Ph.D., Assistant Research Professor whose specialty is aging neurodegenerative disorders such as Parkinson’s, Alzheimer’s and Huntington’s Diseases. This study was conducted on rat cells, not human cells, therefore we can only confirm that Greska’s Carbon-60™ is non-toxic to rats brain cells. Samples of neuronal cell culture of rodent (cerebellar granule cells) were exposed to different concentrations of Greska’s Carbon-60™.  After 24 hours, there were no signs of any negative effects in the viability, morphology or cell death process as comparted to the non-treated cells. The test results from KIHA of Greska’s Carbon-60™ being non-toxic was exciting news! Dr. Paredes continued testing and found other unexpected and interesting results. In his continued testing Dr. Paredes was interested in taking the toxicity experiment to the next level. Given the reported antioxidant properties of Carbon 60, Dr. Paredes investigate the potential of Greska’s Carbon-60™ on cell protection upon confronting the cells with an extremely oxidant environment. In the next set of experiments, the goal was to observe Greska’s Carbon-60™ reaction when hydrogen peroxide (H2O2) was added to the cell culture media to induce cell death. Hydrogen peroxide is a strong oxidizing agent and can be used for bleaching, as a disinfectant and even as a component in rocket fuel. In the scientific research community, it is common practice that hydrogen peroxide is used to induce cell death in scientific research and experiments. In the research article, Is Hydrogen Peroxide a Suitable Apoptosis Inducer for All Cell Types?, it states,   “Hydrogen peroxide is currently the most widely used apoptosis inducer due to its broad cytotoxic efficacy against nearly all cell types.”  Jinmei Xiang, Chunyun Wan, Rui Guo, and Dingzong Guo, “Is Hydrogen Peroxide a Suitable Apoptosis Inducer for All Cell Types?,” BioMed Research International, vol. 2016, Article ID 7343965, 6 pages, 2016. https://doi.org/10.1155/2016/7343965

HOW WAS THE TESTING CONDUCTED?

Using the same rat brain cell culture as was used in the toxicity testing, a control sample and Greska’s Carbon-60™ was prepared. After the addition of hydrogen peroxide, following standard procedure, the expectation is to see significant cell death within 24 hours. When both samples were evaluated after 24 hours, the control sample (non-Greska’s Carbon-60™) showed 80% of cells dead. What was astounding was the brain cell culture with Greska’s Carbon-60™ added, when checked at the 24 hour point, there was only 40% cell death reported. That is an increase in survival rate of 50% when compared to the non-treated cells. According to Dr. Paredes, “Adding hydrogen peroxide to this cell culture is like throwing an atomic bomb in the petri dish. There’s nothing that can prevent the cell death. To see this level of survival rate is remarkable.”  Dr. Paredes continues, “Even though this experiment was done in rodent cells, there is a great probability that such protective effect of the Greska’s Carbon-60™ that we observed, also happens in a whole organism, including humans, this results merit further research to fully explore the potential of this molecule (Greska’s Carbon-60™) in the treatment of devastating conditions where oxidative stress is at the core of cell death”

SO WHY DID GRESKA’S CARBON-60™ SLOW THE CELL DEATH CAUSED BY THE HYDROGEN PEROXIDE?

Before we get to further explanation of the test results, let’s review these definitions so that we can better understand how free radicals, oxidative stress, reactive oxygen species (ROS) and antioxidants fit together.

Free radicals

• Are naturally occurring due to metabolism (exercise)
• May also be caused by UV radiation, tobacco smoke, environmental pollution, toxic chemicals, infections, or poor diet
• Have a missing electron
• Are unstable and highly reactive (constantly looking for an electron to repair itself, causing damage to the neighboring molecule causing a chain reaction or domino effect
• Can damage cells, tissue and DNA through oxidation (oxidative stress)

Reactive Oxygen Species (ROS)

• ROS are signaling molecules
• Body’s natural defenses to detoxify or repair damage
• Environmental stress (UV or heat) can cause ROS to increase
• ROS increase can cause significant cell damage to lipids, proteins and DNA
• Hydrogen peroxide is considered ROS

Hydrogen Peroxide

• Kills cells by starting chemical reaction via lipid peroxidation
• Free radicals “steal” electrons from the lipids in cell membranes

Oxidative Stress

• Is an imbalance between free radicals and antioxidants
• Occurs when there is an excess of free radicals
• Cumulative cell damage caused by excessive ROS,
• Oxidation is a chemical process that produces free radicals

Antioxidant

• Molecules or compounds that inhibit oxidation by donating an electron to a free radical without becoming unstable themselves
• Can be a food, naturally occurring, or dietary supplement
• Support our immune system by attacking foreign bodies

So, we ask the question again—why did Greska’s Carbon-60™ slow down the cell death caused by the hydrogen peroxide?  A simple explanation is, it seems as if Greska’s Carbon-60™ is acting as an antioxidant. Let’s look at the chemical composition for hydrogen peroxide. Hydrogen peroxide is H2O2. That is, 2 atoms of hydrogen and 2 atoms of oxygen. If you remember from your basic chemistry lesson, water is H2O, which is 2 hydrogen atoms and 1 oxygen atom. Hydrogen peroxide is highly toxic to cells, causing cell death. It would appear that Greska’s Carbon-60™ was able to act as an antioxidant and neutralize one of the destructive oxygen atoms, thus dampening the toxicity of hydrogen peroxide. This is a significant and noteworthy finding that deserves further testing and research. Now for the added bonus: Greska’s Carbon-60™ is so small that makes Greska’s C60 an incredibly effective antioxidant.

PART TWO OF TESTING GRESKA’S CARBON-60™ FOR SAFETY, WAS A TEST TO DETECT ANY TOXIC HEAVY METALS

As the second step in confirming Greska’s Carbon-60™ is non toxic and safe, we tested for heavy metals at Steep Hill Laboratories. Steep Hill Laboratories, with many locations throughout the United States is a highly prestigious science and technology company focusing on lab testing, licensing, genetics and research and development for the cannabis industry. Greska’s Carbon-60™ Heavy Metals Test Results We are pleased to report that the Steep Hill Certificate of Analysis (COA) for Greska’s Carbon-60™ shows that our product contains none if these heavy metals:  Arsenic, Cadmium, Lead or Mercury.

WHAT DO: NOT DETECTED (ND), LIMIT OF DETECTION (LOD), AND BELOW LIMIT OF QUANTIFICATION (LOQ) REALLY MEAN?

Not Detected (ND) means the substance being tested for was not found Limit of Detection (LOD) is a term used in analytical chemistry to define the lowest quantity of any substance that can be detected with any analytical procedure. Limit of Quantification (LOQ) is the smallest concentration of a substance that can be reliably measured by an analytical procedure. So to review, the Steep hills laboratory report shows that no heavy metals were found in Greska’s Carbon-60™. In this installment of “The Truth Continues: Testing Greska’s Carbon-60™ for Safety” we will share the results of safety testing conducted at the Knoebel Institute for Healthy Aging (KIHA), University of Denver and  Steep Hill Laboratories in Honolulu, Hawaii. There have been many scientific and research studies conducted regarding the toxicity of Carbon 60 and it has been determined that isolated C60 by itself is not toxic. In fact, the Baati rat study published in 2012 was initially intended as a toxicity test. What the scientists found was unexpected! Not only was C60 is not toxic to rats but it appeared to extend the lifespan of the rats that were given C60. We know Greska’s Carbon-60™ was safe for human consumption, as we are 100% certain our proprietary non-solvent method of producing our C60 uses NO SOLVENTS at any step of the manufacturing process. We then add our C60 to high quality food grade organic sunflower oil. In order to provide the best quality product to our customers, our objective was to verify and provide documentation to our customers that Greska’s Carbon-60™ was not toxic. We wanted confirmation, that not only was Greska’s Carbon-60™ manufacturing process non-toxic but there were no other toxic substances in our product.

PART ONE FOR TESTING GRESKA’S CARBON-60™ FOR TOXICITY

Part One of testing of Greska’s Carbon-60™ for toxicity was conducted at the Knoebel Institute for Healthy Aging (KIHA), University of Denver. Located at the base of the Rocky Mountains in Denver, Colorado, KIHA was founded in 2010 when DU received a generous gift from Betty Knoebel. Mrs. Knoebel was the widow of Ferdinand “Fritz” Knoebel, who was a pioneer in the food services industry. KIHA’s mission is to create and implement solutions for aging issues through multidisciplinary research, education and outreach. Greska’s Carbon-60™ testing was led by Daniel Paredes, Ph.D., Assistant Research Professor whose specialty is aging neurodegenerative disorders such as Parkinson’s, Alzheimer’s and Huntington’s Diseases. This study was conducted on rat cells, not human cells, therefore we can only confirm that Greska’s Carbon-60™ is non-toxic to rats brain cells. Samples of neuronal cell culture of rodent (cerebellar granule cells) were exposed to different concentrations of Greska’s Carbon-60™.  After 24 hours, there were no signs of any negative effects in the viability, morphology or cell death process as comparted to the non-treated cells.