WO2020002387A1

WO2020002387A1 - Composition comprising thymol and hydrogenated palm oil

Info

Publication number: WO2020002387A1
Application number: PCT/EP2019/066923
Authority: WO
Inventors: Elger Funda; Ivan de Jesus GAYTAN PEREZ
Original assignee: Dsm Ip Assets B.V.
Priority date: 2018-06-26
Filing date: 2019-06-26
Publication date: 2020-01-02

Abstract

The present invention relates to a composition comprising hydrogenated palm oil and thymol in a weight ratio from 10:1 to 1:1. Particles comprising or consisting of such composition enable a temperature responsive delivery of thymol. Delivery of thymol is initiated at about body temperature of a broiler and increases flock uniformity.

Description

Composition comprising thymol and hydrogenated palm oil

Technical field The present invention relates to a composition suitable for the oral administration of thymol and its use in the feed industry.

Background of the invention

For centuries, thymol has been used in traditional medicine. It has been shown to possess various pharmacological properties including antioxidant, free radical scavenging, anti-inflammatory, analgesic, antispasmodic, antibacterial, antifungal, antiseptic and antitumor activities (Meeran et al. Pharmacological Properties and Molecular Mechanisms of Thymol: Prospects for Its Therapeutic Potential and Pharmaceutical Development. Front Pharmacol. 2017; 8: 380).

The presence of thymol has been evidenced in the stomach, intestine, and urine after its oral administration with sesame oil at a dose around 500 mg in rats and 1-3 g in rabbits (Schroder V. et al. The excretion of thymol, carvacrol, eugenol and guiacol and the distribution of these substances in the organism. Arch. Exp. Pathol. Pharmak. 1932; 168 331-353).

Thymol is a white crystalline substance with a melting point ranging from 49°C to 51 °C. Therefore, thymol crystals as such could be added to a premix comprising vitamins, minerals and other additives. However, thymol as such has an unpleasant taste and smell which makes it less palatable (Nieddu M. et al. Improvement of thymol properties by complexation with cyclodextrins: In vitro and in vivo studies. Carbohyd. Polym. 2014; 102 393-399).

Robbins encapsulated thymol in gelatine capsules before administering it to dogs (Robbins B. H. Quantitative studies on the absorption and excretion of certain resorcinols and cresols in dogs and man. J. Pharmacol. Exp. Therapeut. 1934; 52 54-60). Whereas such an approach reduces the smell and taste of thymol, it is not possible to add gelatine capsules to feed premixes. Gelatine capsules are by far too expensive for the use in the feed industry. Furthermore, a premix containing two-piece gelatine capsules would easily de-mix, i.e. it would have a very poor blend uniformity.

There is a need for a cost-effective manner to reduce smell and/or taste of thymol by providing a suitable galenic formulation of thymol. The sought-after formulation must release of thymol once the animal has swallowed the formulation, should be storage stable, should be suitable for providing premixes with high blend uniformity, should be easy to manufacture, must be non-toxic and must fulfill the applicable regulatory requirements.

There is also a need for a manner to improve flock uniformity. In commercial practice, flock uniformity is important. Many wholesale purchasers of chicken meat insist on a narrow weight range. Failure to meet these specifications can incur severe economic losses (cf. Madsen, T.G. and Pedersen, J.R. (2010) Broiler Flock Uniformity. Feedstuffs 82: 12-13).

Summary of the invention

The problems underlying the present invention are solved by encapsulating thymol in a matrix which releases thymol at body temperature.

Temperature responsive delivery of thymol is challenging. The matrix of such advanced delivery format must be non-toxic, must fulfill regulatory

requirements, must be cheap, must be available at large quantities and should initiate the release of thymol at about body temperature. The present invention relates to a composition comprising hydrogenated palm oil and thymol. When hydrogenated palm oil starts to become soft, the release of thymol is initiated. At the melting point of the composition, thymol is fully released.

Softening of the material can be seen in a Differential Scanning Calorimetry (DSC) curve as a“shoulder” of the melting point. Softening point denotes the onset temperature of this shoulder, respectively the lowest temperature at which softening starts.

Hydrogenated palm oil without thymol melts over a relatively broad range, due to the presence of a and b polymorphic forms. In the experiments of the inventors, hydrogenate palm oil without thymol did not fully melt until a temperature of 53°C had been reached. At body temperature, there was only very little softening of the hydrogenated palm oil.

Surprisingly, the addition of thymol to hydrogenated palm oil lowers the softening point of hydrogenated palm oil. At body temperature, there is already substantial softening of such mixture. The temperature at which thymol is released from the mixture can be fine-tuned by using an appropriate weight ratio between hydrogenated palm oil and thymol.

If desired, the release of thymol can be further enhanced by the addition of eugenol: a composition that comprises hydrogenated palm oil and thymol is softer at body temperature when said composition also comprises eugenol.

A preferred embodiment of the invention relates to particles which comprise or consist of the composition of the invention. Such particles are preferably obtained by spray chilling encapsulation. Spray chilling encapsulation is an easy and cheap manufacturing method. The particles of the invention are storage stable because thymol is encased in an matrix of hydrogenated palm oil. Despite of said encasement, the particles of the invention release thymol once body temperature has been reached.

Surprisingly, flock uniformity is improved when particles which comprise or consist of the composition of the invention are fed to birds (e.g. broilers) over a period of preferably at least 20 days, more preferably over a period of at least 35 days, starting with day-old birds. Such particles can be fed as such. Alternatively, a premix comprising such particles can be fed. Preferably, feed comprising such particles and/or comprising such premix is fed to birds over a period of at least 20 days, more preferably over a period of at least 35 days, starting with day-old broilers. Therefore, the present invention also relates to the use of particles according to the invention, to the use of the premix of the invention, and to the use of feed of the invention to increase flock uniformity. Preferably, the use relates to the uniformity of a flock of broilers.

Detailed description of the invention The composition of the invention comprises hydrogenated palm oil, thymol and optionally piperine. Preferably, the composition of the invention comprises hydrogenated palm oil and thymol.

At room temperature, hydrogenated palm oil is solid. Hydrogenated palm oil may have one or multiple (e.g. two) peaks in its DSC melting curve.

Preferably, the hydrogenated palm oil of the invention has a maximum in its DSC curve between 40°C and 60°C, preferably between 42°C and 55°C and most preferably between 45°C and 50°C, wherein said maximum indicates an endothermic process (melting, i.e. said maximum may be a negative peak). In one embodiment, the lowest melting point of the hydrogenated palm oil of the invention is from 39°C to 45°C and is preferably from 40°C to 44°C. The hydrogenated palm oil of the invention may be partially hydrogenated palm oil, fully hydrogenated palm oil or a mixture thereof. Most preferably, the hydrogenated palm oil of the invention is fully hydrogenated palm oil.

At body temperature, hydrogenated palm oil is softer when thymol and/or eugenol has been added. Advantage can be taken of this effect such that the release of thymol is triggered and/or enhanced at about body temperature.

Advanced delivery formats have been known for some time. Known is in particular the use of temperature-sensitive polymers for drug delivery.

The concept of the present invention is different: instead of using an expensive, non-GRAS temperature-sensitive polymer, a surprisingly simple and thus cost-effective composition is used which releases, depending on the temperature, more or less thymol.

At room temperature (25°C), the composition of the invention is solid and encapsulates thymol very effectively. At body temperature of a broiler (40°C), however, the composition becomes soft and the encapsulation of thymol is no longer as effective as before: the release of thymol is initiated. Thus, the present invention also relates to the use of the composition of the invention for controlling the release of thymol by temperature.

Body temperature induced release of thymol may then be enhanced by a mechanical effect. Without wishing to be bound by theory, it has been hypothesized that soft, deformable compositions are deformed in the crop and/or gizzard of the broiler. Said deformation results in an increased surface-to-volume ratio which is expected to further enhance the release of thymol from the composition of the invention. The composition of the invention comprises hydrogenated palm oil and thymol in a weight ratio from 10:1 to 1 :1 . The release of thymol at body temperature can be fine-tuned by modifying said weight ratio. In a preferred embodiment of the invention, the weight ratio between hydrogenated palm oil and thymol is from 8:1 to 2:1 , is preferably from 7:1 to 3:1 and is most preferably from 6:1 to 5:1.

The release of thymol at body temperature can be further enhanced by adding eugenol. Thus, a preferred embodiment of the invention relates to a composition comprising hydrogenated palm oil, thymol and eugenol, wherein the weight ratio between hydrogenated palm oil and thymol is from 10:1 to 1 :1 , preferably from 8:1 to 2:1 , more preferably from 7:1 to 3:1 and most preferably from 6:1 to 5:1.

The composition of the invention may have any shape. However, the composition of the invention is preferably a powder which comprises preferably spherical particles. The size of the particles should be adapted to the intended use. In case of a feed additive for broiler, the particles must be small enough to be captured by a chicken.

Any known method can be used to shape the composition of the invention. However, a particularly useful method for encapsulating thymol with hydrogenated palm oil is spray chilling encapsulation. For such method, it may be beneficial to add at least one auxiliary compound. Preferably, said at least one auxiliary compound is silicic acid, calcium sulfate, stearic acid, monopropylene glycol and/or starch. When the composition of the invention comprises eugenol and/or said at least one additive, the composition comprises preferably 5-30 weight-% thymol, more preferably 10-20 weight-% thymol and most preferably 12-18 weight-% thymol, based on the total weight of the composition. In one embodiment, the composition of the invention comprises eugenol, thymol, piperine and at least one additive. In a preferred embodiment, the composition of the invention consists of eugenol, thymol, piperine and at least one additive.

The person skilled in the art is familiar with spray chilling encapsulation. Such method comprises preferably the steps:

i. providing a mixture comprising molten hydrogenated palm oil,

thymol, optionally eugenol and optionally at least on auxiliary compound, said molten hydrogenated palm oil being preferably fully hydrogenated palm oil;

ii. cooling the mixture provided in step i) by spraying said mixture into a cooling medium, said cooling medium being preferably air.

Thus, the present invention also relates to a method of encapsulating thymol by spray chilling, wherein the composition of the invention is molten before being sprayed.

Spray chilling encapsulation improves storage stability of volatile compounds. This also applies to the particles of the invention which are, in addition to having improved storage stability, capable of releasing of thymol once body temperature has been reached.

The composition or particles of the invention may be added to a premix comprising vitamins, minerals and/or other additives. Thus, the present invention also relates to a premix comprising the composition and/or the particles of the invention. Premixes comprising the particles of the invention hardly de-mix, i.e. they have a good blend uniformity.

Typically, the premix of the invention is added to food or feed. Thus, the present invention also relates to food or feed comprising the premix, the composition and/or the particles of the invention. Preferably, the feed of the invention comprises 0.5-2 weight-%, more preferably 1 weight-% premix of the invention, based on the total weight of the feed. Other preferred amounts are indicated in below table:

The present invention also relates to the use of the particles of the present invention for increasing flock uniformity. It also relates to the use of the premix of the present invention for increasing flock uniformity. And it also relates to the use of the feed of the present invention for increasing flock uniformity.

In the context of the present invention, the term“flock” refers preferably to a flock of birds, more preferably to a flock of broilers.

The person skilled in the art is familiar with the concept of flock uniformity. Flock Uniformity (FU) can be expressed as the coefficient of variation (CV) in body weight. Increased CV values are synonymous with decreased uniformity, i.e. a wider spread in body weights above and below the flock average (of. Hughes et al. Flock Uniformity - Is it important and how is it assessed? Aust. Poult. Sci. Symp. 2017). In the context of the present invention, the average final body weight (FBW) of a bird in a specified pen (e.g. pen number 1 ) is used as basis for the calculation.

Typically, a pen comprises at least 10 or at least 15 broilers. Because broilers within one pen share one source of feed, the feed intake of an individual broiler within the pen cannot be determined. What can be determined is the total feed intake of all broilers within the specified pen. Due to this reason, the final body weight (FBW) of each and every individual broiler is not measured during standardized trials. What is measured is the total final body weight of all broilers in the specified pen. When said total final body weight of all broilers is divided by the number of broilers within said pen, the average final body weight (FBW) of a broiler in the specified pen is obtained. The term

“FBWd35/pen” refers to the average final body weight (FBW) of a broiler in a specified pen when the respective broilers have been fed for 35 days.

Preferably, said 35 feeding days start with day-old broilers. For further details of the calculation, reference is made to example 3.

Figures

FIGURE 1 shows the DSC melting curve of the samples of example 1. The following symbols are used

On the x-axis, the temperature is shown in °C. On the y-axis, energy flow is shown (normalized, i.e. Watt/g of the composition). Negative energy flow indicates endothermic processes (e.g. melting). Positive energy flow corresponds to exothermic processes. Examples

Example 1

Four samples were prepared by the following process: 1. Melting of hydrogenated palm oil in a 70°C water bath.

2. Addition of the other optional ingredients, one after the other while stirring at 200 rpm.

3. Speed up of stirring (500 rpm) and mix for 3 min.

4. Cool down slowly at room temperature. Temperature was set to 70°C and controlled during steps 1-3 of the preparation process. All ingredients are commercially available. Thymol (purity: 99%) was purchased at VWR Chemicals, eugenol (purity: 99%) at Merk KGaA. Fully hydrogenated palm oil is commercially available.

The composition of samples 1 to 4 is shown in below table:

Table 1

For each of the samples, a melting curve was measured by Differential Scanning Calorimetry, using a Discovery DSC (TA Instruments, Waters GmbH, Eschborn). The obtained DSC melting curves are shown in Figure 1. Melting points were determined by the extrapolated peak onset temperatures (2nd heating cycle at 5°C per minute from -10 °C to 90 °C). Eugenol, if present, was not included as it remained always in liquid state.

Sample 1 (hydrogenated palm oil only) shows two peaks, corresponding to a and b polymorphic forms. Sample 1 was not completely molten until a temperature of 51.3 °C had been reached.

The analysis of sample 2 (thymol only) revealed a melting point of 48.7°C and thus, confirmed the accuracy of the method used in example 1.

The partial replacement of hydrogenated palm oil with thymol (sample 3) had a major effect on the melting point: the melting point of sample 3 was determined as 43.9°C. At 38°C the heat flow difference between sample 1 and sample 3 was -0.12588 W/g. At the body temperature of an average broiler (40°C), said difference was even more pronounced. DSC melting curve of sample 3 indicates that softening of sample 3 starts at an even lower temperature. In the crop of broiler, said softening will trigger a deformation of the product. As a result of the increased surface-to-volume ratio of the deformed product, the release of thymol is expected to be further enhanced.

Softening and melting point are even lower if the composition comprises eugenol in addition to thymol (sample 4). The compositions of sample 3 and sample 4 enable a temperature responsive delivery of thymol, which is further enhanced by mechanical effects in the crop and/or gizzard of a chicken.

Example 2

Particles comprising the composition of the invention were produced as follows:

Molten hydrogenated palm oil was mixed with thymol, eugenol and selected auxiliary compounds. To obtain particles, the liquid mixture was cooled by spraying (spray chilling encapsulation). Organoleptic inspection of the obtained particles confirmed a reduced smell. The thus obtained particles were used to prepare a premix. Feed comprising 1 weight-% of said premix, based on the total weight of the feed, was then fed to broiler.

Example 3

The effect of the particles comprising the composition of the invention on Flock Uniformity (FU) was tested as follows:

A trial was run in one room with multiple pens. Day-old Cobb500 male broilers were randomly allocated to each pen (25 broilers/pen). Three different poultry feeds were tested. For each kind of poultry feed, 12 pens were allocated. Thus, for each kind of poultry feed, 300 broilers were used (12 pens ^* 25 broilers/pen = 300 broilers) were used. To test all three different poultry feeds, a total of 900 boilers (3 types of feed ^* 300 broilers/feed = 900 broilers) was used. For each of the three different poultry feeds, standard deviation (SD) and the coefficient of variation (CV) was calculated and expressed as follows:

CV =SD (FBWd35/pen) /Mean (FBWd35/pen)^*100

Increased CV values are synonymous with decreased flock uniformity (FU). For an overview of the experimental setting and the details of the statistical analysis, see Table 2.

Table 2 The result of the test is shown in Table 3.

Table 3

Example 3 clearly shows that flock uniformity is improved (i.e. lower CV value) when raising broilers with the feed of the invention (poultry feed 1 ). Flock uniformity decreases (i.e. higher CV value) if the feed comprises thymol and eugenol as such (i.e. without hydrogenated palm oil; poultry feed 2) or if the feed comprises none of these three compounds (poultry feed 3; negative control).

Example 4

Example 3 was repeated at a different research center under similar conditions with a different kind of broilers:

The trial was run in a room with multiple pens. Day-old Ross308 male broilers were randomly allocated to each pen (16 broilers/pen). Three different poultry feeds were tested. For each kind of poultry feed, 12 pens were used. Thus, Thus, for each kind of poultry feed, 192 broilers were used (12 pens ^* 16 broilers/pen = 192 broilers) were used. To test all three different poultry feeds, a total of 576 boilers (3 types of feed ^* 192 broilers/feed = 576 broilers) was used.

The result of the trial of example 4 is shown in Table 4.

Table 4

Example 4 confirms the outcome of example 3: flock uniformity is clearly improved (i.e. lower CV value) when raising broilers with the feed of the invention. Flock uniformity decreases (i.e. higher CV value) if the feed comprises thymol and eugenol as such (i.e. without hydrogenated palm oil; poultry feed 2) or if the feed comprises none of these three compounds (poultry feed 3; negative control).

Claims

1. Composition comprising hydrogenated palm oil and thymol, wherein the weight ratio between hydrogenated palm oil and thymol is from 10:1 to 1 :1.

2. Composition according to claim 1 , wherein the weight ratio between hydrogenated palm oil and thymol is from 8:1 to 2:1 , is preferably from 7:1 to 3:1 and is most preferably from 6:1 to 5:1 and/or wherein said hydrogenated palm oil is fully hydrogenated palm oil.

3. Composition according to claim 1 or 2, wherein said composition further comprises eugenol.

4. Composition according to any one of claims 1 to 3, wherein said

composition further comprises at least one auxiliary compound, and wherein said at least one auxiliary compound is preferably silicic acid, calcium sulfate, stearic acid, monopropylene glycol and/or starch.

5. Composition according to claim 3 or 4, wherein said composition

comprises 5-30 weight-% thymol, preferably 10-20 weight-% thymol and most preferably 12-18 weight-% thymol, based on the total weight of the composition.

6. Particles comprising or consisting of the composition according to any one of claims 1 to 5.

7. Particles according to claim 6, wherein said particles are obtainable by a method comprising the steps

i. providing a mixture comprising molten hydrogenated palm oil, thymol, optionally eugenol and optionally at least on auxiliary compound;

ii. cooling the mixture provided in step i) by spraying said mixture into a cooling medium.

8. Particles according to claim 6 or 7, wherein said particles enable a temperature responsive delivery of thymol.

9. Premix comprising the composition according to any one of claims 1 to 5 or comprising the particles according to any one of claims 6 to 8.

10. Premix according to claim 9, wherein one kilogram of said premix

comprises 0.1 g to 10 g of the composition according to any one of claims 1 to 5, or wherein one kilogram of said premix comprises 0.1 g to 10 g of the particles according to any one of claims 6 to 8.

1 1. Food or feed, comprising the premix of claim 9 or claim 10.

12. Feed according to claim 1 1 , wherein one ton of said feed comprises 10 g to 100 g of the composition according to any one of claims 1 to 5, or wherein one ton of said feed comprises 10 g to 100 g of the particles according to claim 7 or 8.

13. Method of encapsulating thymol by spray chilling, wherein the

composition according to any one of claims 1 to 5 is molten before being sprayed.

14. Use of particles according to any one of claims 6 to 8, or use of the

premix of claim 9 or claim 10, or use of the feed of claim 1 1 or 12 to increase flock uniformity.

15. Use according to claim 14, wherein the uniformity of a flock of birds is increased, and wherein said flock of birds is preferably a flock of broilers.

16. Use of a composition comprising hydrogenated palm oil and thymol for controlling the release of thymol by temperature.

17. Use according to claim 16, wherein the composition according to any one of claims 1 to 5 is used.