WO2002052950A1

WO2002052950A1 - Organomineral adsorbent of mycotoxin as an animal feed additive, procedure for production and application

Info

Publication number: WO2002052950A1
Application number: PCT/YU2001/000032
Authority: WO
Inventors: Magdalena Tomasevic-Canovic; Milutin Dumic; Olivera Vukicevic; Aleksandra Dakovic; Sinisa Milosevic; Djordje Avakumovic; Isidor Rajic
Original assignee: Institut Za Tehnologiju Nuklearnih I Drugih Mineralnih Sirovina
Priority date: 2000-12-28
Filing date: 2001-12-24
Publication date: 2002-07-11
Also published as: HUP0300377A3; HRPK20020706B3; HUP0300377A2; SK12362002A3; CZ20022905A3; HRP20020706A2; BG107098A

Abstract

Organomineral adsorbent of mycotoxins as an animal feed additive, procedure for production and application, is obtained by known processes, technological preparation of zeolitic tuff, containing more than 80% of mineral clinoptilotite/heulandite, three steps comminution, wet classification and water suspended. Suspension 20-50 wt% of zeolitic tuff is mixed in a mixer on 4 000 r/m with addition of organic component -quaternary amine with long carbon chain, the most suitable are dioctadecyldimethyl amine, octadecylthreemethyl amine, octadecyldimethylbenzyl amine and similar compounds, in the amount of 0.5-5 meq/100g of zeolitic tuff. Suspension is filtered, dryed at temperature 80-100°C, desintegrated up to the particle size below 100 micrometers and than packed in powdered state. Adsorbent of mycotoxins possess balanced surface charge and hydrophobicity level and in accordance to mentioned a new active sites, which absorb mycotoxins of different polarities. Organomineral adsorbent of mycotoxins is added to the animal feed directly or through premixes in the amount of 0.2 wt% to feedmix.

Description

Organomineral adsorbent of mycotoxins as an animal feed additive, procedure for production and application

Field of technic invention concerned

Organomineral mycotoxins adsorbent as animal feed additive, procedure for production and application belongs to the animal feed, addition of mineral additives. According to the International clasification of Patents (MKD⁶), Section A, Sign mark A23K1/175.

Technical problem

Technical problem, which is solved by this invention, consist of: how to obtain by chemicotechnological preparation of zeolitic tuff, containing over 85% mineral clinoptilolite/heulandite and strictly controled amount of certain organic compound (quaternary amine long carbon chains), surface modified asorbent, in order to attain controled surface charge and hydrophobicity level, what have to lead to organomineral adsorbent, as animal feef additive, which, in the same time adsorbs mycotoxins of different properties in digestive tract of animals.

The state of technic

New aproach on mycotoxins effects prevention, e.g. feed detoxification polluted by mycotoxins, in the World, are focused on efficient adsorbent development, which are added to to the animal feed. Their task is to effectively, selectively and very fast adsorb mycotoxins in situ, in digestive tract, when mineral adsorbent together with polluted feed enter the tract. Directly, in the electrolyte of gastric juice, adsorbent bind mycotoxins and in such a way decreases their bioavaillability, e. g. blocked their resorption in organs and cancel their toxicity.

Mycotoxins, from chemical side, represent very wide spectar of complex organic compounds with different functional groups and therefore different characterictics. According to its use in the case coexist, more than one mycotoxin with different characteristics (polarity, functional groups, anionic or cationic character etc), adsorbent need to have in the same time active centers on the surface, different but strictly controled characteristics. As this problem is very complex, investigations in this field, in the World, are very intensive, particularly last years.

On the World market, one can find, more new products for mycotoxins adsorption usually based on modified clay minerals (HSCAS, Novosil, Myco-ad - USA; Mycofix - Plus, Biomin - Austria; Mycosil - Mexico). By modification of clay minerals, surface characteristics were changed and in such a way improvent and enlargement of adsorption efficiency for wide spectar of mycotoxins was obtained. Mineral adsorbent of mycotoxins is known, as animal feed additive, having trade name Min-a-Zel, obtained in accordance to YU Patent No 48589, assignee Institute for Technology of Nuclear and Other Mineral Raw Materials, Belgrade, Yugoslavia. The Patent understand chemical-technological processing of zeolite tuff, containing 85% of clinoptilolite, by three steps comminition, activation and milling with sodium chloride up to the particle size max. 60 micrometers. Obtained powdered product is a mineral adsorbent of mycotoxins out of animal feed, in the digestive tract of animals. It is added to the mycotoxin contaminated feed in the amount of 0.2 to 0.5%, depending on the contaminated level of feed mix.

It is known, also, mycotoxins mineral adsorbent, improved characteristics, as an animal feed additive, obtained by procedure according to the announced Patent YU No P- 177/98 (assignee Institute for Technology of Nuclear and Other Mineral Raw Materials, Belgrade, Yugoslavia). Patent contain procedure for production of mineral adsorbents applaying three steps comminution of zeolitic tuff, containing 85% clinoptilolite, up to particle size 150 micrometers, than with potassium chloride, in dependance of the exchangable potassium content in the raw tuff, it is activated and milled up to max. 60 micrometers. Obtained material is wet classified on centrifuge, separating particles below 10 micrometers and packed as a 25% suspension in water for future application. If the 25% suspension is filtrated, dried and desintegrated, than mycotoxin mineral adsorbent is obtained as a powder. As an animal feed additive, it is mixed in the feed mix in the amount of 0.3 to 0.5%.

Both mycotoxin mineral adsorbent, in accordance to Patent YU No 488589 and Yu No P- 177/98 have shown as a good mycotoxin adsorbents, for moleculs belonging to the group of polar (aflatoxins and others) over 90%, while mycotoxins with lower polarity (zearalenone, ochratoxin A, T-2 toxin)were adsorbed 30-50%).

Exposing the essence of invention

Mycotoxins, secondary metabolites produced by fungi (moulds) are highly toxic for numerous number of animals (swines, cattle, sheeps, horses, dogs, cats, rabbits), poultry (chickens, turkey, ducks, gooses), laboratory animals and humans. Mycotoxicosis are deseases caused by consuming feedstuff contaminated by mycotoxins, they are not infective but could not be treated by any medication. Well known mycotoxicosis of domestic animals are aflatoxicosis, ochratoxicosis, zearalenone toxicosis and trichothecene toxicosis. On the World level, estimation is that more than 25% of cereals are contaminated by mycotoxins. As the resorption of mycotoxins is very fast in meat, milk and eggs, their presence in animal foodstuff is toxic, not only for animals but also for humans.

The essence of invention lies in controled correction of surface properties charge by modification processes with organic compounds - radicals, on previously processed zeolitic tuff by known procedures - three steps comminution, wet classification to 25%) suspension.

That is achived by addition of controled amount of chosen quaternary amine with long chains in the suspension of clinoptilolite/heulandite tuff of certain concentration, enabling formation of organomineral complex, having different characteristics comparing to starting materials (surface charge, hydrophobicity level etc). Zeolitic tuff, containing more than 80%) of mineral clinoptilolite/heulandite, prepared by known technological processes comminution, wet classification and preparation of 25%> suspension, attaining balance of exchangeable cations, maintaining negative surface charge as a result of isomorphic supstitution of Al³⁺ by divalent cations (Ca or Mg²⁺). Such a surface adsorbs polar mycotoxins. Further organic modification of zeolitic tuff influence the exchange of surface charge and changes of adsorption level for less polar mycotoxins, especially for zzearalenone (96%>), ochratoxin A (96%) and T-2 toxin (85%). In the same time, adsorption level for aflatoxins (99%) does not decreases. As far as for ergot alcaloids, which adsorbs on unorganic form over 80% but on organo modified analog over 90%>.

That could be explaned in a such a way: obtained organomineral adsorbent of mycotoxins — organozeolite, as boundary phase had not only hydrophillic negative surface charge, but it is transfered partially in hydrophobic, having the consequence in better adsorption of less polar mycotoxin moleculs. That, in the same time, leads to partial exchange of the surface charge, from negative to positive, what is controlled by the amount of added organic compound used for modification. Positive charge on the surface are now a new active centers for adsorption of polar moleculs, too, having anionic character.

Presence of organic phase, used for modification, is limited not only for obtaining desired product characteristics (surface charge, hydrophobicity) but, also, from the side of permited amounts to be consumed by a live organism, because over the certain concentration they become toxic.

Description of the invention, in details

A new organomineral adsorbent of mycotoxins of controled surface charge and controled hydrophobicity level, according to this invention, is obtained as a product of reaction take place in suspension, by known processes, previously prepared and processed mineral component - zeolitic tuff, containing > 80% mineral clinoptilolite/heulandite and quaternary amine high molecule weight. Organomineral adsorbent of mycotoxins - organozeolite as a result of ion exchange reaction between inorganic cations on the zeolitic tuff surface (Ca²⁺, Mg²⁺, Na⁺, K⁺) and long chain quaternary amine such as dioctadecyldimethyl amine, octadecylthreemethyl amine, octadecyldimethylbenzyl amine or similar amines. Quaternaiy amine is exchanged quantitatively and compitly irreversible with inorganic cations on the external surface of the zeolitic tuff. Molecule of the quaternary amine is very large to be settled in structural channels of zeolitic tuff and interior exchange sites stay free for reaction with smaller cations, such as inorganic cations.

I Characteristics of mineral component - zeolitic tuff

1) Particle size distribution:

100% below 150 micrometers

80% below 63 micrometers

50% below 20 micrometers

Chemical composition

SiO₂ 63-68 wt.%

Al₂O₃ l l-14 wt.%

Fe₂O₃ 0.8-2,5 wt.%

MnO 0.01-0.03 wt.%

CaO 2.5-4.5 wt.%

MgO 0.8-1.5 wt.%

Na₂O 0.8-1.5 wt.%

K₂O 1.0-2.0 wt.%

L.I. 10.5-14.5 wt.% 3) Heavy metals

Pb max 30 ppm

Cr max 10 ppm

Sb max 20 ppm

Cd max 5 ppm

4) Cation exchange capacity 160 ± lO meq/lOOg

5) Acid stability (by dissolved Al) at pH 3 max 50 ppm Al at pH 2.5 max 100 ppm Al

6) Accesory minerals

Any traces of erionite is not permited.

Undesirable presence of fibre minerals: azbestos, sepiolite.

II Organic component

Organic additive e.g. organic compound is prepared as a dispersion in warm distiled water. Organic component is quaternary amine with long carbon chains, usually, dioctadecyldimethyl amine, octadecylthreemethyl amine, octadecyldimethylbenzyl amine or similar compound are used.

Organic component is added in amount of 0.5 - 5.0 meq/lOOg of prepared mineral component - zeolitic tuff, containing min. 80% mineral clinoptilolite/heulandite. Type of ammine as far as its amount depends on type and quantity of mycotoxins to be adsorbed. Organomineral adsorbent is prepared in accordance to the next example:

Example 1

In the procedure for production of this organomineral adsorbent of mycotoxins, zeolitic tuff containing min 80% of mineral clinoptilolite/heulandite, by known processes, as three steps comminution up to max 100 micrometers, wet classifying and prepared as water suspension of 20-50 wt% is transfered in the mixer and mixed (min 4 000 r/m) or better in turbo mixer.After that, activator - organic component in the amount of 0.5-5.0 meq/lOOg mineral component, dispersed in hot distiled water is added.. Ratio solid : liquid is from 1 : 1 to 1 : 5.

Organic component is quaternary amins with long carbon chain, usually dioctadecyldimethyl amine, octadecylthreemethyl amine, octadecyldimethylbenzyl amine or similar ones. Amine type and quantity to be applicated depends on type and quantity of mycotoxins to be adsorbed. Depending on the content of zeolitic tuff, after the reaction is completed, the obtained suspension is filtered (20%) and cake is dried directly (50%) in batch dryirs at the temperature from 80-100°C. After drying the cake is desintegrated in "knife mill" up to particle size below 100 micrometers. Obtained product is organomineral adsorbent of mycotoxins, as powder which is packed for the appication.

Such organomineral adsorbent of mycotoxins, obtained by organic modification, well balanced surgace charge is achieved, which enables wide spectmm of mycotoxin, of different polarity, adsorption. Amine is bind to zeolitic tuff by cation exchange of natural exchangeable cations (Ca²⁺) which is located at the surface of zeolitic tuff. Having the balance of exchangeable cations content, organomineral adsorbent of mycotoxins, is of controlled surface charge and controlled hydrophobicity level. Organomineral adsorbent of mycotoxins, which is hydrophobic, can be easily mixed in various mixtures, such as premixes or feedmixes.

Organomineral adsorbent of mycotoxins under conditions in vitro is adsorbing without desorption. the next mycotoxins:

99% aflatoxin Bl 94% zearalenone 96% ochratoxin A 82% T-2 toxin 50% DAS 96%o ergosine 92% ergocornine 98%> ergocristine 98% ergocriptine 97%) ergotamine

Obtained organomineral adsorbent of mycotoxins does not adsorb vitamins (A, D, E, group B), aminoacids and does not influence on availability of mikroelements (Cu, Zn, Mn and Co); does not adsorb P, N and Ca.

In application, obtained organomineral adsorbent of mycotoxins, in powder state, is added to animal feed directly or through premix, in amount of 0.2 wt% to feed mix to preventing toxication by mycotoxins present in animal feed, without any harmful effects. The amount of 0.2 wt% of this mycotoxin adsorbent is sufficient to prevent toxication of animals, which consumed the feed contaminated with zearalenone, up to 10 mg/kg. In the same time, the presence of 0.2 wt% of this adsorbent in feed mix has no negative effect on state of health and production results and could be used in prevention. This should be pointed out, for the reason, if animals are poisoned by mycotoxins, then there is no way to detoxicate the animals neither by adsorbent nor by medicaments. Organomineral adsorbent of mycotoxins prevent toxication only if it is present in the feedstuff polluted with mycotoxins. This organomineral adsorbent will bind mycotoxins in digestive tract of animals before they would be resorbed by organism, because the adsorption rate of mycotoxins on adsorbent is higher than rate of resorption, even in the case of zearalenone, for which is showed that the 50% of initialy amount is resopted for less than 5 minute, while 99% for less than 1 hour. In order to prevent toxication by mycotoxins, it is necessery that adsorbent to be present in animal feed, as a preventive additive, because in case of animal toxication, e. g. resorption of mycotoxins in organism, detoxification is such a cases is imposible. Later addition of organomineral adsorbent would prevent only further toxication of animals.

Example 2 (application in vivo)

Results of application effects of the mycotoxins organomineral adsorbent were executed on three pig farms on piglets (5-25 kg):

I Farm experimental group - number of pigs 123 and control group 115 pigs;

II Farm — both, control and experimental group - 200 pigs and

III Farm 35 pigs for both groups.

In Farms I and II, experimental group of pigs were fed with addition of 0.2%> of organomineral adsorbent and in Farm III 0.1% of adsorbent was added. Control group were fed with same feed but without adsorbent. The feed contained 5-7 mg/kg of zearalenone. All other conditions were identical.

Obtained results show:

1) Presence of 0.2%) of organomineral adsorbent had a great influence on pigs state of health: number of sick pigs in experimental groups were considerable lower than in control groups (Farm I, less for 7%, Farm II - less for 40%). - Number of dead pigs were lower in experimental than in control groups (Farm I - less for 7%, Farm II - less for 66%>).

2) Addition of 0.1% (Farm III) and more 0.2% (Farms I and II) of organomineral adsorbent in animal feed had influence on production parameters:

Pigs fed with the feed containing 0.2% of mineral adsorbent had a higher gain per feeding day for 10% (Farm I), 4.4%> for Farm II and 10.7% for Farm III (0.1% of adsorbent).

Feed conversion is lower for 14%> (Farm I), 4% for Farm II and 2% for Farm III in experimental group.

Example 3 (application in vivo)

Investigations executed on fattering swine (25-100 kg) had shown that addition of 0.1% or 0.2% of organomineral adsorbent of mycotoxins in the feed, containing 5.5 mg/kg of zearalenone.

Farm I, both experimental and control group had 70 swine each. For experimental group

0.1% of adsorbent was added to the feed. Feedstuff contains 5.2 mg/kg of zearalenone.

Farm II 0.2% of adsorbent was added for experimental group (122 fattering swine) and control group had 114 swine. Feedstuff contains 5.5 mg/kg of zearalenone.

In Farm I, fattering swine of experimental group had higher gain per feeding day for

2.36% comparing to control group. Feed conversion in experimental group was lower for 2.6%. In Farm II the gain was higher for 3% and in the same time feed conversion was lower for 7%.

Claims

Patent Claims

1. Organomineral adsorbent of mycotoxins as an animal feed additive, obtained by technological processing of previously prepared zeolitic tuff, which is inert to aminoacids, vitamins, microelements, nonresorptive in organism, point out that in mineral component - zeolitic tuff, containing minimum 80% of mineral clinoptilolite/heulandite, contain organic component from the group of quaternary amines with long carbon chain, in the amount of 0.5 to 5 meq/lOOg of mineral component - zeolitic tuff, having the next characteristics: chemical composition - SiO₂ 63-68 wt.%; Al₂O₃ 11-14 wt.%; Fe₂0₃0.8-2,5 wt.%; MnO 0.01-0.03 wt.%;CaO 2.5-4.5 wt.%; MgO 0.8-1.5 wt.%; Na₂O 0.8-1.5 wt.%; K₂0 1.0-2.0 wt.%; loss of ignition 10.5-14.5 wt.%; heavy metals: Pb max 30 ppm, Cr max 10 ppm, Sb max 20 ppm, Cd max 5 ppm; cation exchange capacity: 160 ± 10 meq/lOOg; acid stability expressed by disolved Al: at pH 3 max 50 ppm Al, at pH 2.5 max 100 ppm Al; particle size below 100 micrometers, with amine bind to the mineral component - zeolitic tuff by cation exchange with naturaly exchangeable cations (usually Ca²⁺) which are located on the external surface of zeolitic tuff; in such a way organomineral adsorbent has controled surface charge and controled hydrophobicity level what make it as a very efficient adsorbent of mycotoxins of different polarities.

2. Procedure for production organomineral adsorbent of mycotoxins, according to Demand 1, by which, known procedures, zeolitic tuff formerly prepared and processed by three steps comminution, wet classified and water suspended, point out as mineral component - zeolitic tuff, containing over 80%) of mineral clinoptilolite/heulandite, particle size below 100 micrometers, in water suspension 20-50 wt% of solid phase mix in reactor with mixer on 4 000 r/m with addition of organic component, presented by quaternary amine with long carbon chain, such as dioctadecyldimethyl amine, octadecylthreemethyl amine, octadecyldimethylbenzyl amine and similar compounds; in the amount of 0.5-5 meq/lOOg of mineral component - zeolitic tuff, depending on type and quantity of mycotoxins to be adsorbed; organic component is previously disperged in warm distiled water, solid : liquid ratio 1 : 1 to 1 : 5; obtained suspension is filtered (20%); cake is dried (50% is directly dried) in batch dryer at temperature 80-100°C up to the humidity 10%; after drying cake is desintegrated to the particle size below 100 micrometers; than packed in powdered state as organomineral adsorbent of mycotoxins characteristics according to Demand 1.

3. Application of organomineral adsorbent of mycotoxins of different characteristics, having balanced exchangeable cation content, controlled surface charge and hydrophobicity level, with wide spectra of mycotoxin adsorption, is in addition in powdered state to animal feed, directly or by premixes, in the amount of 0.2 wt% to feedmix, preventing toxication of animals by mycotoxins, present in animal feedstuff, without harmfull effects.