patented technology – mycotoxin binder. • speed – allows

Patented Technology – Mycotoxin Binder

• Are toxic secondary metabolites produced by several fungi, particularly by species of: Aspergillus Fusarium Penicillium Claviceps Alternaria.

• It has been estimated that at least 300 of these fungal metabolites are potentially toxic to animals and humans

• MYCOTOXINS CANNOT BE CURED, ONLY PREVENTED!

MYCOTOXINS (I)

• The most common mycotoxins are: aflatoxin B1 (AFB1) ochratoxin A (OTA) zearalenone (ZEA) deoxynivalenol (DON, “vomitoxin”) T-2 and HT-2 toxins fumonisins (FUM)

MYCOTOXINS (II)

• Mycotoxins are produced by fungi during growth, handling and storage of agricultural commodities.

• Their global occurrence is considered to be a major risk factor.

• Worldwide, approximately 25% ofcrops are affected by mycotoxins.

• The economic consequences of mycotoxin contamination areprofound, and often crops with large amounts of mycotoxin have to be destroyed.

MYCOTOXINS (III)

• There is an increasing awareness of the hazards posed to both human and animal health by the presence of toxins produced by fungi in food and feed.

• Mycotoxins have a diversity of chemical structures which accounts for different biological effects: Carcinogenic Mutagenic Teratogenic Oestrogenic Neurotoxic Immunotoxic, etc.

MYCOTOXINS (IV)

• In farm animals, mycotoxins can cause, among others: decreased performance feed refusal poor feed conversion diminished body weight gain immune suppression reproductive disorders residues in animal food products

MYCOTOXINS (V)

• The best way to reduce mycotoxins from contaminated feed is the use of mycotoxin binders.

• The aim of these additives is to inhibit the uptake of mycotoxins by an animal in vivo.

• Mycotoxin binders act like a ‘chemical sponge’ and adsorb mycotoxins in the gastrointestinal tract, thus preventing the uptake and distribution to target organs.

MYCOTOXIN BINDERS (I)

• The efficacy of the adsorption depends on the chemical structure of both the binder and the mycotoxin.

• The most important feature for adsorption is the physical structure of the binder:– total charge – charge distribution– size of the pores– accessible surface area

• On the other hand, the properties of the adsorbed mycotoxins, like polarity, solubility, shape and charge distribution, also play a significant role.

MYCOTOXIN BINDERS (II)

• Commonly used mycotoxin binders are: Inorganic binders:

Activated Carbon Mineral based binders (Zeolites, Bentonite, Caolin, etc.)

Ogranic binders: Esterified glucomannan (Saccharomyces cerevisiae) Resins (cholestyramine, etc.) Enzymes (eubacterium, etc.)

MYCOTOXIN BINDERS (III)

• Most commonly used are: Bentonites Zeolites Caolins Others

MINERAL BASED BINDERS

• The most common natural mineral zeolites are: Analcime Chabazite Clinoptilolite Heulandite Natrolite Phillipsite Stilbite

ZEOLITES

• STABILITY!

• SELECTIVITY!

• EFFICACY!

WHY PATENT CO. USES CLINOPTILOLITE?

• Clinoptilolite is an alumosilicate mineral of a volcanic or sedimentary origin.

• According to the classification of the European Commission (Regulation (EC) No 1831/2003), and the registry of additives (EU Register of Feed Additives), clinoptilolite is classified into group 1 - Technological additives (codes E567 and E568).

CLINOPTILOLITE

• Three-dimensional lattice structure of the mineral, with easily replaceable positive ions - cations (Na+, K+, Mg2+, Ca2+).

• This makes surface area available for mycotoxins binding!

CLINOPTILOLITE STRUCTURE

• pH stability - Unlike other aluminosilicate minerals, clinoptilolite is pH stable, retaining its mineral structure in all pH (from 1 to 10).

• This makes it stable in the stomach and intestines, thus favorable for use as animal feed supplement.

CLINOPTILOLITE IS STABLE!

Stomach is very acidicpH 2-3

Small IntestinepH 7

Large IntestinepH 8

• Selectivity - Because of its lattice structure, clinoptilolite shows selectivity during adsorption.

• It adsorbs only mycotoxins, while leaving vitamins, minerals, amino acids in the feed!

• This is not the case with some other minerals (bentonite).

CLINOPTILOLITE IS SELECTIVE!

• Clinoptilolite is effective in binding polar mycotoxins!• In animals, Clinoptilolite has limited beneficial effect

against zearalenone, fumonisin B1, ochratoxin A, and trichothecenes (deoxynivalenol, T-2 toxin)

• In order to improve this characteristic Patent co. hasdeveloped a special patented technological process of clinoptilolite surface modification, therefore improving its affinity for adsorption of all mycotoxins.

CLINOPTILOLITE BINDING EFFICACY

THIS PRODUCT IS CALLED

• Minazel Plus is patented under EU Patent No. 1363854

MINAZEL PLUS – PATENTED TECHNOLOGY (I)

1. Raw material reception:a) Quality Control:

I. HumidityII. Chemical composition,III. Clinoptilolite contentIV. Heavy metal contentV. Dioxine contentVI. Granulometry

b) Raw material preparation:I. Heat treatmentII. GrindingIII. Mechanical activation

MINAZEL PLUS – PATENTED TECHNOLOGY (II)

2. Trybochemical surface modification:a) Organic cation addition to

clinoptiloliteb) Physico-Chemical process – partial

change of the Clinoprilolite surface polarity by attaching long chained cations.

c) Physico-Chemical process requires:I. Three-step comminution procedure II. High kinetic energyIII. Controled temperature

MINAZEL PLUS – PATENTED TECHNOLOGY (III)

3. Quality Control:a) Capacity of Cationic

Exchangeb) Hydrophobicity of the

productc) IN VITRO testing of

mycotoxin adsorption efficacy

MINAZEL PLUS – PATENTED TECHNOLOGY (IV)

• 20 years of experience in mycotoxicosis problem solving

• Product generations:1. Minazel - 19932. Minazel Plus – 20003. NEW generation mycotoxin

binder – 2012

PATENT CO. AND MYCOTOXINS

Na1+ Ca2+K1+ Na1+ Ca2+ Ca2+ Na1+ Ca2+ K1+ Na1+ Mg2+ Na1+ Na1+ Ca2+ Na1+

K1+

NATURAL CLINOPTILOLITE SURFACE (Negative surface charge is caused by substitution of Si4+ with Al3+ or Fe3+)

Easily replaced Cations (K, Ca, Na, Mg)

CHAINS OF ORGANIC CATION(activation)

POLARMYCOTOX

POLARMYCOTOX

POLARMYCOTOX

POLARMYCOTOX

POLARMYCOTOX

POLAR MYCOTOXIN BINDING (Afla, Ergo...)

NON POLARMYCOTOX NON POLAR

MYCOTOX

NON POLARMYCOTOX

NON POLARMYCOTOX

NON POLARMYCOTOX

NON POLARMYCOTOX

NON POLAR MYCOTOXINSBINDING (Zea, Ochra, T-2...)

MINAZEL PLUS (I)

• Adsorption of less polar mycotoxins requirespartial correction of the mineral surfacecharge and surface hydrophobicity.

• This is achieved by using long-chain organiccations.

• Physical and chemical features of the outersurface of minerals are dramatically changedby adding long-chain organic cation

• This results in development of twofold layerof organic ligandos to which non-polarmycotoxins are being adsorbed.

Activated clinoptilolite

• Hydrophobic surface is a boundary link of theorganocomplex, which makes it compatiblewith non-polar organic mycotoxins’ molecules(zearalenonee, ohratoxins, T-2 toxin and alike).

• New, active centres, which are formed on thesurface, ensures efficient binding of thisadsorbent (over 90%) not only with aflatoxinsand ergot alkaloids but also with zearalenone,ohratoxin A, T-2 toxin and other mycotoxins aswell.

• This makes MINAZEL PLUS more powerful andeffective in comparison to other products.

MINAZEL PLUS (II)

Activated clinoptilolite

• Stability – Minazel Plus is not absorbed in the body• Does not contain damaging components (heavy

metals and unfavorable minerals)• Does not adsorb vitamins, amino acids and

microelements.• Speed – allows it to adsorb mycotoxin in the

stomach, before they show negative effect in theintestine.

• Dosage – 1-2kg/ton of feed

PRODUCT CHARACTERISTICS (I)

Minazel Plus is safe for the environment:• Bonds between Minazel Plus and

mycotoxins are very stable, and show no signs of weakening over time.

• Mycotoxins can not be released in the soil, and re-enter the food chain.

PRODUCT CHARACTERISTICS (II)

Surfactant modified zeolites new efficient adsorbents for mycotoxinsMicroporous and Mesoporous Materials 61 (2003) 173–180

Magdalena Tomasevic-Čanovic a, Aleksandra Dakovic a,*,George Rottinghaus b, Sr dan Matijasevic a, Mirjana Đuricic aa Institute for Technology of Nuclear and Other Mineral Raw Materials, P.O. Box 390, 11000 Belgrade, Yugoslaviab College Veterinary Medicine, University of Missouri, Columbia, MO 65211, USA

MINAZEL PLUS IN VITRO ADSORPTION

Aim of the study:

1. To test IN VITRO mycotoxin adsorption of modified clinoptilolite Minazel Plus

2. To test IN VITRO chemical stability after an electrolyte treatment at pH 1, 7 and 10


Materials and methods (I)• Clinoptilolite – characteristics check:

– Particles size <100 μm– Total cation exchange capacity (CEC) = 145 meq/100 g– Calcium was the dominant ion in an exchangeable

position.


Materials and methods (II)• Stability check:

– The samples (5 g of Minazel Plus) were placed into 100 ml electrolyte solution (pH 1, 7 and 10), shaken at 20oC for 60 min, filtrated and the Minazel Plus was dried at 80oC.

– Samples were heated (20–800oC) in an air atmosphere, with a heating rate of 10oC/min.

– Results were obtained using IR spectrometry.


Materials and methods (III)• Adsorption check:

– For mycotoxin adsorption studies 100 mg of Minazel Plus sample was added to 10 ml of pH 3 buffered solution, containing either:

• aflatoxin B1 (2ppm)• zearalenone (2 ppm)• ochratoxin A (2 ppm)• ergopeptine alkaloids (0.5 ppm)

– After 30 minutes HPLC analyses were performed in order to measure % of adsorption



Results (I)• IR spectra of Minazel Plus, before

and after treatment at pH 1 (6/1), pH 7 (6/7) and pH 10 (6/10) arepresented in Fig. 1

• There is no difference between the IR spectra, which suggests that Minazel Plus is stable over a pH range of 1–10.

Results (II)• HPLC analyses were

performed• Summary were presentaed

in the table


Mycotoxin Adsorption (%)

Aflatoxin B1 99%

Zearalenone 94%

Ochratoxin A 96%

Fumonisin B1 86%

T-2 Toxin 83%

Ergot Alkaloids 97%

Adsorption effects of Minazel Plus -adsorption speed of Zearalenone-

Aleksandra Daković*, Srđan Matijašević*, George E. Rottinghaus**

*Institure for Technology of Nuclear Other Mineral Raw Materials, Belgrade** Veterinary Medical Diagnostic Laboratory, College of Veterinary Medicine, Missouri, USA

1) Zearalenone test solutions (1.000 ppm) for adsorption studies were prepared and adjusted to pH 3, 7 and 9.

2) 15 ml screw cup tubes, along with 6 mg of Minazel Plus were added in buffer.

3) Tubes were placed on a rotate shaker at room temperature.4) Tubes were centrifugated after at 13.000 rpm for 5 minutes, and HPLC

analzsis was performed to investigate % of adsorption.

MATERIALS AND METHODS

Speed of adsortion was tested by calculation of rezidual zearalenone content in a solution after 5 min, 30 min and 60 min of contact. Results are shown in a table.

TRIAL RESULTS

Contact time zearalenone / Minazel Plus % of adsorption

5 minutes 51.51 %

30 minutes 76.17%

60 minutes 93.04%

Adsorption effects of Minazel Plus adsorption behavior in the presence

of Aminoacids and VitaminsMagdalena Tomašević-Čanović*, M. Dumić*, Olivera Vukićević*, Mirjana Đuričić* and S. Jovanović**

*Institure for Technology of Nuclear Other Mineral Raw Materials, Belgrade** Institute Vinča, Vinča

Aminoacid adsorption:1) Tryptophan and phenylalanine were used at 1mM/dm3 solutions in

electrolyte (16.5 mg of phenylalanine/100 ml and 20.6 mg tryptophan/100 ml)

2) Test method for determination: 1 g of Minazel Plus was added to the solution, and the suspension was placed in the water bath at 37°C.

3) After 2 hours concentration of non adsorbed phenylalanine and tryptophan were determined by HPLC.


Results of the analysis has shown that investigated ligands (tryptophan and phenylalanine) were not adsorbed. In figure 1 the spectra of tryptophan in clear electrolyte (Figure 1-1) and in the presence of Minazel Plus (Figure 1-2) are presented.IT IS CLEAR THAT ALL TRYPTOPHAN WAS PRESENT IN THE ELECTROLYTE AFTER CONTACT WITH MINAZEL PLUS FOR 2 HOURS. IT MEANS THAT MINAZEL PLUS DID NOT BIND ANY AMINOACIDS!

TRIAL RESULTS

Clear system Presence of Minazel Plus

In figure 2 the spectra of phenylalanineare presented, showing the same behavior – NO ADSORPTION. No difference in the intensity in the clean solution and in the supernatant after 2 hours Minazel Plus / phenylalanine contact was noted!

TRIAL RESULTS


Vitamin adsorption1) Vitamin A, D3 and E were used in following amounts:

• 1.5 mg of Vitamin A/100 ml of electrolyte• 12.5 mg of Vitamin D3/100 ml of electrolyte• 10.0 mg of Vitamin E/100 ml of electrolyte

pH was adjusted to 7.0 with 0.1 N NaOH.2) Test method for determination: 1 g of Minazel Plus was added to the

solution, and the suspension was placed in the water bath at 37°C. 3) After 2 hours concentration of non adsorbed vitamin A, D3 and E were

determined by HPLC.


Results of the analysis has shown that vitamin A, vitamin D3 and vitamin E were not adsorbed. In figure 3 curves obtained for vitamin A in electrolyte (Curve 1) and in supernatant after 2 hours Minazel Plus/vitamin A contact (Curve 2) are given.THE SAME INTENSITY CONFIRMS THAT THE PROCESS OF ADSORPTION DID NOT OCCUR!

TRIAL RESULTS


The results obtained for Vitamin E (Figure 5) also shows that Minazel Plus does not bind vitamin E!

TRIAL RESULTS

Presence of Minazel Plus

Clear system

patented technology – mycotoxin binder. • speed – allows

Documents