SUPPLEMTING COLLOIDAL IRON

INTELLECTUAL PROPERTY OF DVS BioLife LtdCONFIDENTIAL

NOT FOR PUBLIC CIRCULATION

FECOL

Iron deficiency is still a problem in animal and human nutrition, especially in the earlier stages of life, during gestation, and in parasite infestations (Church et al., 2003). One of the most bioavailable sources of this element is in the form of ferrous sulfate(McDowell, 1997), but it has the inconvenience of giving food a metallic taste, and accelerating.

Parenteral iron therapy is today widely used for the treatment of irondeficiency anemia. Patients with chronic kidney disease (CKD) also frequently need treatment with parenteral iron preparations in addition to erythropoietin stimulating agents [1]. For renal failure patients on dialysis, the average iron requirements due to blood loss are equivalent to 1–3 g of elemental iron per year [2]. This can easily be accomplished by frequent low dose IV iron administrations, during the regular dialysis sessions. From initial, generalized use innephrology parenteral iron therapy has spread in recent years to otherdisease areas; gastroenterology [3], cardiology [4,5], oncology [6], pre/post operatively [7], obstetrics’, and gynecology [8]. However, care providers in these segments have less frequent patient contact,

resulting in an increased demand for convenient administration of large IV iron doses in one clinical session.(http://www.staff.uni-mainz.de/kolb/pdf_dateien/132_isomaltoside_FeO_science.pdf)

What Is Anemia?Anemia is a condition that develops when your blood lacks enough healthy red blood cells or hemoglobin. Hemoglobin is a main part of red blood cells and binds oxygen. If you have too few or abnormal red blood cells, or your hemoglobin is abnormal or low, the cells in your body will not get enough oxygen. Symptoms of anemia -- like fatigue -- occur because organs aren't getting what they need to function properly.

Anemia is the most common blood condition in the U.S. It affects about 3.5 million Americans. Women and people with chronic diseases are at increased risk of anemia. Important factors to remember are:Certain forms of anemia are hereditary and infants may be affected from the time of birth.

Women in the childbearing years are particularly susceptible to iron-deficiency anemia because of the blood loss from menstruation and the increased blood supply demands during pregnancy.

Older adults also may have a greater risk of developing anemia because of poordiet and other medical conditions.

There are many types of anemia. All are very different in their causes and treatments. Iron-deficiency anemia, the most common type, is very treatable with diet changes and iron supplements. Some forms of anemia -- like the anemia that develops during pregnancy -- are even considered normal. However, some types of anemia may present lifelong health problems.

What Causes Anemia?There are more than 400 types of anemia, which are divided into three groups:1. Anemia caused by blood loss2. Anemia caused by decreased or faulty red blood cell production3. Anemia caused by destruction of red blood cells

Anemia Caused by Blood LossRed blood cells can be lost through bleeding, which can occur slowly over a long period of time, and can often go undetected. This kind of chronic bleeding commonly results from the following:

Gastrointestinal conditions such as ulcers, hemorrhoids, gastritis(inflammation of the stomach), and cancer

Use of nonsteroidal anti-inflammatory drugs (NSAIDs) such asaspirinor ibuprofen, which can cause ulcers and gastritisMenstruation and childbirth in women, especially if menstrual bleeding is excessive and if there are multiple pregnancies

Anemia Caused by Decreased or Faulty Red Blood Cell ProductionWith this type of anemia, the body may produce too few blood cells or the blood cells may not function correctly. In either case, anemia can result. Redblood cells may be faulty or decreased due to abnormal red blood cells or a lack of minerals and vitamins needed for red blood cells to work properly. Conditions associated with these causes of anemia include the following:1. Sickle cell anemia2. Iron-deficiency anemia3. Vitamin deficiency4. Bone marrow and stem cell problems5. Other health conditionsSickle cell anemia is an inherited disorder that affects African-Americans. Red blood cells become crescent-shaped because of a genetic defect. They breakdown rapidly, so oxygen does not get to the body's organs, causing anemia. Thecrescent-shaped red blood can cells also get stuck in tiny blood vessels, causing pain.Iron-deficiency anemia occurs because of a lack of the mineral iron in the body. Bone marrow in the center of the bone needs iron to make hemoglobin, thepart of the red blood cell that transports oxygen to the body's organs. Without adequate iron, the body cannot produce enough hemoglobin for red bloodcells. The result is iron-deficiency anemia. This type of anemia can be causedby:

An iron-poor diet, especially in infants, children, teens, vegans, and vegetariansThe metabolic demands of pregnancy and breastfeeding that deplete a woman's iron storesMenstruationFrequent blood donationEndurance trainingDigestive conditions such as Crohn's disease or surgical removal of part of the stomach or small intestineCertain drugs, foods, and caffeinated drinksVitamin-deficiency anemia may occur when vitamin B12 and folate are deficient. These two vitamins are needed to make red blood cells. Conditions leading to anemia caused by vitamin deficiency include:

1. Megaloblastic anemia: Vitamin B12 or folate or both are deficient2. Pernicious anemia: Poor vitamin B12 absorption caused by conditions such as

Crohn's disease, an intestinal parasite infection, surgical removal of partof the stomach or intestine, or infection with HIV

Dietary deficiency: Eating little or no meat may cause a lack of vitamin B12, while overcooking or eating too few vegetables may cause a folate deficiency.

Other causes of vitamin deficiency: pregnancy, certain medications, alcohol abuse, intestinal diseases such as tropical sprue and celiacdisease

During early pregnancy, sufficient folic acidcan help prevent the fetus from developing neural tube defects such as spina bifida.Bone marrow and stem cell problems may prevent the body from producing enough redblood cells. Some of the stem cells found in bone marrow develop into red blood cells. If stem cells are too few, defective, or replaced by other cells such as metastatic cancer cells, anemia may result. Anemia resulting from bone marrow or stem cell problems include: Aplastic anemia occurs when there's a marked reduction in the number of stem

cells or absence of these cells. Aplastic anemia can be inherited, can occur without apparent cause, or can occur when the bone marrow is injured by medications, radiation, chemotherapy, or infection.

Thalassemia occurs when the red cells can't mature and grow properly. Thalassemia is an inherited condition that typically affects people of Mediterranean, African, Middle Eastern, and Southeast Asian descent. This condition can range in severity from mild to life-threatening; the most severeform is called Cooley's anemia.

Lead exposure is toxic to the bone marrow, leading to fewer red blood cells. Lead poisoning occurs in adults from work-related exposure and in children whoeat paint chips, for example. Improperly glazed pottery can also taint food and liquids with lead.

Anemia associated with other conditions usually occur when there are too few hormones necessary for red blood cell production. Conditions causing this typeof anemia include the following:

Advanced kidney disease Hypothyroidism Other chronic diseases, such as cancer, infection, lupus, diabetes, and

rheumatoid arthritis Old age

Anemia Caused by Destruction of Red Blood CellsWhen red blood cells are fragile and cannot withstand the routine stressof thecirculatory system, they may rupture prematurely, causing hemolytic anemia. Hemolytic anemia can be present at birth or develop later. Sometimes there is no known cause. Known causes of hemolytic anemia may include:

Inherited conditions, such as sickle cell anemia and thalassemia Stressors such as infections, drugs, snake or spider venom, or certain foods Toxins from advanced liver or kidney disease Inappropriate attack by the immune system (called hemolytic disease of the

newborn when it occurs in the fetus of a pregnant woman) Vascular grafts, prosthetic heart valves, tumors, severe burns, chemical

exposure, severe hypertension, and clotting disorders In rare cases, an enlarged spleen can trap red blood cells and destroy them

before their circulating time is up.

(http://www.webmd.com/a-to-z-guides/understanding-anemia-basics? )

TRADITIONAL TREATMENTS:Iron deficiency anemia treatmentThis usually involves taking iron supplements to replace the lack of intake of iron in diet or excess loss of iron.The most commonly prescribed supplement is ferrous sulphate. It is taken as pills two or three times a day.Oral iron preparations come with a host of side effects that include nausea, vomiting, abdominal pain, heartburn, constipation, diarrhea, black stool and blackening of teeth, gums and tongue.Another alternative is ferrous gluconateVitamin C supplements helps absorb iron better

Vitamin B12 deficiency anaemia treatmentThis can be treated by taking vitamin B12.Methylcobalamin is preferable.Vegetarians or vegans are generally prone for the deficiency of this.

Anemia due to folate deficiencyFor folate deficiency anemia daily folic acid tablets are prescribed.Folate tablets are usually prescribed along with Vitamin B12 supplements. This is because folic acid treatment can sometimes improve the symptoms masking an underlying vitamin B12 deficiency.If a vitamin B12 deficiency is not detected and treated at this stage there could be severe damage to the brain, nerves and spinal cord due to vitamin B12 deficiency.Folate is found in broccoli, green cabbage, wheatgerm, pulses, nuts, green leafy vegetables etc.

Treatment for severe anemiaWhen the anaemia is more severe, a blood transfusion is often necessary.

Sickle cell anemia treatmentPatients with sickle cell anemia need a healthy diet, supplements of folic acid, vitamin D and zinc and avoid triggers for crises.This includes smoking, alcohol, overexertion, dehydration, cold and hot temperatures, constricting clothes etc.There is no cure for sickle cell anaemia, but the frequency and severity of crises and their complications can be reduced. They need complete vaccinations against flu, pneumococcus meningitis, Hepatitis B and other diseases to prevent infections.

Anemia due to infectionAnemia caused by an infection will usually improve when the infection is treated. This is especially true for newborns with severe infections called sepsis.

Treatment for an enlarged spleenIn some forms of haemolytic anemia there may be an enlarged spleen.The spleen may be surgically removed to prevent RBCs from being removed from circulation or destroyed too rapidly.

Anemia in pregnancyIf the hemoglobin concentration is less than 9.0 g per dL anemia in pregnancy is diagnosed.Anemia is managed with oral dose of 60 to 120 mg per day of iron. Patient is evaluated after four weeks of therapy.

Anemia and bone marrow treatmentSome medications are prescribed to stimulate the bone marrow to produce more RBCs. This is useful in aplastic anemia and leukemias.Bone marrow transplantation may also be used. In this procedure, bone marrow cells taken from a matching donor (usually with a genetic matche.g. a sibling or blood relation).This is then injected into the vein. This then travels through the bloodstream to the bone marrow and produced new blood cells.(http://www.news-medical.net/health/Treatment-of-anemia.aspx)

Antagonistic nature of the Minerals.

Copper-Iron InteractionRuminants consuming forage based diets can be exposed to high levels of iron. Grazing animals can consume high levels of iron as a consequence of soil consumption during grazing and through consumptionof soil contaminated forages. Suttle (1975) estimated that during winter, soil ingestion can exceed 10% of the dry matter intake of grazing sheep and cattle. This level of soil ingestion markedly reduced copper absorption (Suttle, 1975). High dietary iron has been shown to reduce copper status in cattle (Standish et al., 1971; Campbell et al., 1974; Humphries et al., 1983; Phillipo et al., 1987; Mullis et al., 2003) and sheep (Prabowo et al., 1988; Grace and Lee, 1990). It is believed that sulfide in the rumen combines with iron to form ferrous sulfide complexes that dissociate in the low pH abomasum where the sulfide then forms insoluble complexes with copper (Gengelbach et al., 1994).

Over sixty years ago a German scientist named Abderhalden conducted a series of experiments comparing how several species absorbed differentforms of iron. He found that animals fed with food poor in iron, plus in addition of inorganic iron, were unable in the long run to produce as much hemoglobin as those, receiving a natural iron-sufficient diet.

While the inorganic iron may be absorbed into the body, it is not utilized in the formation of hemoglobin, but remains unused within thetissues. Abderhalden also concluded that any apparent benefit of the inorganic iron resulted from its stimulating effect.

Chemically, it is true that iron in the bloodstream and iron in nails are the same and that calcium in rocks (known as dolomite) is identical to calcium in the bones.

However, it is a grave error to believe that the body can digest and assimilate and utilize powdered nails and crushed rocks.

Mineral Sources of Iron:Ferrous sulfate heptahydrate FeSO4(H2O)7 20% Fe

Ferrous Oxide FeO Ferric Oxide Fe2O3 69.94% Fe

FeO(OH)) or hydrated (FeO(OH)·nH2O) forms.

The monohydrate (FeO(OH)·H2O) might otherwise be described as iron(III) hydroxide (Fe(OH)3), and is also known as hydrated iron oxide or yellow iron oxide.Iron(II) Oxide (FeO)Colour: BlackMolecular Weight: 71.84

Iron (II,III) Oxide (Fe3O4) AKA MagnetiteColour: BlackMolecular Weight: 231.53

Iron(III) Oxide (Fe2O3) AKA Hematite, RustColour: Red-brownMolecular Weight: 159.69

Iron(IV) Oxide (FeO2)Colour: RedMolecular Weight: 159.70

Extent of absorption (%) 7-10%

Ferric compounds are much less well absorbed when HCL secretion in thestomach is low compared to ferrous compound.

Dietary iron availability and absorption is usually depressed by high dietary intakes of phosphate, calcium, phytates, copper and zinc.

Reducing substances such as vitamin C enhance the absorption of non-haem iron.

TOXICITY OF RED/ YELLOW IRON OXIDE-Iron oxide is not toxic, although some dangers are present when the fumes or dust of iron oxide are inhaled.-Inhalation of any metallic fumes or dust can cause irritation to the throat and nasal area. They can also irritate the eyes.-Iron oxide in high concentrations in the form of dust or fumes can cause metal fume fever.-Metal fume fever is not extremely critical to health, causing a fever, sore/irritated/dry throat and mouth, and a metallic taste. Thisfever usually only lasts from 12 to 48 hours.

-Extended inhalation of iron oxide in high concentrations in the form of dust or fumes can cause pulmonary effects, eventually resulting in siderosis (inflammation of the lungs).

DATA FOR DOSE ARRIVING

Normal Serum Iron Levels

Males (adult) 75 - 175 mcg/dL

Females (adult)

65 - 165 mcg/dL

Children 50 - 120 mcg/dL

 Oral Iron Supplements

Iron SupplementTablet Size Elemental Iron

Usual Dose for Irondeficiency anemia1

Ferrous fumarate 325 mg 106 mg 1 tab bid

Ferrous gluconate 325 mg 36 mg 2 tabs tid

Ferrous sulfate325 mg 65 mg 1 tab tid

Polysaccharide iron

Niferex® caps:Niferex® -150 forte:Niferex® Elixir:

60 mg150 mg100 mg/5 ml

1 cap qd (Forte)

1. Usual total daily dose (IDA): 150-200 mg Fe (elemental iron) per day [Range: 60 to100 mg elemental iron every 12 hours or up to 60 mg elemental iron every 6 hours]

Factors contributing to arriving inorganic iron supplementations are shown below.

mg blood iron mL blood

g hemoglobin

mg iron

————— =————— x————— x—————

lb body weight

lb body weight mL blood

g hemoglobin

a. Blood volume . . . . . . . . . . . . . . . .65 mL/kg of body weight

b. Normal hemoglobin (males and females)over 15 kg (33 lbs) . . . . . . . . . . . .14.8 g/dl15 kg (33 lbs) or less . . . . . . . . . .12.0 g/dl

c. Iron content of hemoglobin . . . . . . . . . . . . . .0.34%d. Hemoglobin deficite. Weight

Based on the above factors, individuals with normal hemoglobin levels will have approximately 33 mg of blood iron per kilogram of body weight (15 mg/lb).

Note: The table and above data are applicable for dosage determinations only in patients with iron deficiency anemia; they are not to be used for dosage determinations in patients requiring iron replacement for blood loss.

Iron-carbohydrate complexes also have significant toxicity, and also they are expensive.

Revolutionary innovative solutions to overcome every of the above deficiencies can effectively be evolved using the aid of nanotechnology.

IRON PROTEINATE is a well known organic mineral compound to be used asa supplemental nutritional source of Iron in swine, dairy, poultry, beef, and pet diets designed to replace the added dietary inorganic elemental Iron.

IMPROVING THIS BY EMBEDDING IRON INSIDE AN AMINO ACID AND ENCAPSULATE IT IS THE NOVELCOCEPT OF OUR FECOL

This Technology Provide i)                   Vastly Enhancing the bioabsorption

ii)                  Vastly Reducing the Toxicity

iii)                 Making the product Bland in taste

iv)                 Improving Solubility

v)                  Making the product safe so as to not

affect/deteriorate the activity of

other nutrients and also not directly interact and will not

cause free radical generation and lipid peroxidation.

Suspended nanoparticles of iron compounds in a colloidal solution ofwater, oil and a surfactant

Nanotechnology adopts the concepts of PHYSICS and not chemistry and thus does not change the chemical structure of the molecule, and can hence be incorporated immediately.

Nanoparticles can be broadly grouped into two: namely organic and inorganic nanoparticles.

Organic nanoparticles may include carbon nanoparticles (fullerenes) while some of the inorganic nanoparticles may include magnetic

nanoparticles, noble metal nanoparticles (like gold and silver) and semiconductor nanoparticles (like titanium dioxide and zinc oxide).

FECOL COMES UNDER ORGANIC MINERAL NANO PARTICLE

“The formulation of micronutrients and nutraceuticals in the design of functional foods brings enormous technological challenges. The incorporation of micronutrients and/or nutraceuticals can compromise the product functionality. Issuesoften encountered are related to unwanted changes in the product physico-chemical stability, appearance, texture, flavour, taste and bioavailability due to inherited instability or interactions with other ingredients. This review intends to present the general strategies in using colloidal dispersions as delivery systems formicronutrients and nutraceuticals. Some illustrative examples will be given on how colloidal delivery systems can be utilised in the design of novel functional foods.

(http://pubs.rsc.org/en/content/articlelanding/2008/sm/b804863k#!divAbstract)

Simple iron salts are considered too toxic for parenteral administration, since ionization of these compounds liberates free iron, and iron is a transition element capable of catalyzing free radical generation and lipid peroxidation (Brown, Moore, Reynafarje & Smith, 1950; Minotti & Aust, 1992). Of the two common valences, Fe(ll)is the most reactive form leading to the production of highly reactivehydroxyl radicals by the Fenton reaction, or alkoxyl and peroxyl radicals from the breakdown of lipid peroxides (reviewed by Gutteridgeand Halliwell, 1990).

Therefore only colloidal iron compounds, that are polynuclear ferric hydroxide carbohydrate complexes, are currently in use for parenteral administration of iron. These compounds are characterized by a complexstructure in which a core of ferric iron lies surrounded by a complex carbohydrate structure, so that iron core is shielded from coming in direct contact with plasma or cells. Examples of these compounds include iron dextran, polymaltose, gluconate, saccharate and chondroitin sulfate.

(http://www.google.com/patents/WO2001000204A1?cl=en)

Cox and coworkers have found that 1-2% of iron present in fresh ampules of Imferon® is ferrous iron, present as an extremely weak ferrous-dextran complex and Presumably, depolymerization of iron-dextran complex releases free dextran molecules (Mol. Wt. ~ 6000 Dalton, ~ 33 glucose units) and ionic iron. (Cox, King & Reynolds, 1965).Thus ionic iron usage is common and well known.

Whereas in our product FECOL Iron is ionized and embedded in a matrix of Colloidal Near Nano Size Amino acid and all this matrix is again

coated with a biopolymer and thus the iron ion is shielded and is not coming into contact directly thus avoiding free radical generation and lipid peroxidation and does not lead to the production of highly reactive hydroxyl radicals by the Fenton reaction, or alkoxyl and peroxyl radicals from the breakdown of lipid peroxides and hence considered safe.

We are of the opinion that ionised iron entrapped in an aminoacid and further shielded by a biopolymer coating can be safely administered bythe parenteral route since this iron is to be tightly complexed to theligand, and thereby causing clinically insignificant release of free iron, and the kinetic properties of this novel double protected iron allows a rapid transfer of iron to plasma transferring. However we are not preferring this route and wish to use it as anoral administration.

FECOL FORTE CONTAINSIron Colloidal 500 ppmMethyl CobalaminFolic acidAscorbic acidCitric acidTartaric acidEclipta alba extractSylybum marianum extractSwertia chirraita Extract2-Methoxy-5-nitrophenol

DISCLAIMERWaiver: The information presented in this publication is based upon the research and experience of DVS BioLife Ltd. The information above is believed to be accurate and represents the best information currently available to us. The information in this MSDS is intended for persons having related technical skills.However, we make no warranty of merchantability or any other warranty, expressor implied, with respect to such information, and we assume no liability resulting from its use. Users should make their own investigations to determine the suitability of theinformation for their particular purposes.

No representation or warranty is made concerning the accuracy or completeness of the information presented in this publication. DVS BioLife Ltd makes no warranty or representation of any kind, express or implied, including without limitation any warranty of merchantability or fitness for any particular purpose, and no warranty or representation shall beimplied by law or otherwise. Any products sold by DVS BioLife Ltd are not warranted as suitable for any particular purpose to the buyer. The suitability of any products for any purpose particular to the buyer is for the buyer to determine. DVS BioLife Ltd shall in no event be liable for any special, incidental, or consequential damages.In no event shall DVS BioLife Ltd be liable for any claims, losses, or damagesof anythird party or for lost profits or any special, indirect, incidental, consequential or exemplary damages, howsoever arising, even if DVS BioLife Ltdhas been advised of the possibility of such damages.The implied warranty of fitness for a particular purpose is limited to the extent the products are used for the purpose or uses described on the product's label or in any written instructions or materials distributed to thebuyer by DVS BioLife Ltd and is hereby disclaimed should buyer use the products in a manner inconsistent with this uses or purposes described therein.