A novel liquid formulation

Abstract

The present invention relates to liquid compositions of ω-nitrooxy-C3-10alkane-1-ols as well the use thereof.

Description

[0001] A Novel Liquid formulation

[0002] The invention relates to liquid compositions of co-nitrooxy-Cs- alkane-l -ols as well the use thereof.

[0003] 3-nitrooxypropanol (3-NOP, also known as 3-nitrooxy-propan-1-ol or 1 ,3-propanediol mononitrate) has been reported to be highly efficient in reducing the formation of methane in ruminants without affecting microbial fermentation in a way that would be detrimental to the host animal (WO-2012-084629-A1 ).

[0004] 3-nitrooxypropanol is mainly used into animal feeds in the form of a 10 % adsorbate on silica while there is an ongoing need for liquid product forms. A stable liquid product form comprising stable 3-nitrooxypropanol would furthermore be highly desirable, as this would also allow the supplementation of ruminants with 3-nitrooxypropanol directly via the drinking water.

[0005] Surprisingly it has now been found, that the addition of 3-nitrooxypropanol into one or more selected liquid solvents provides a stable and clear solution.

[0006] Figure 1 provides photos of the liquid composition of the present application containing 2% (left) and 9% (right) 3-NOP in refined rapeseed oil after 30 days storage at 25°C.

[0007] Figure 2 provides photos of the liquid composition of the present application containing 22% 3-NOP in propane-1 ,2-diol after 9 months storage at 25°C.

[0008] In the first aspect, the present invention provides a liquid composition, which comprises a co-nitrooxyC3-ioalkane-1 -ol dissolved in at least one liquid solvent.

[0009] The term “co-nitrooxyC3-ioalkane-1 -ol” refers to the linear co-nitrooxyalkane-1 -ols having 3 to 10 carbon atoms such as 3-nitrooxypropane-1 -ol (3-NOP, also referred to as 3-nitrooxypropanol), 4-nitrooxybutane-1 -ol, 5-nitrooxypentane-1-ol, 6-nitrooxyhex- ane-1 -ol, 7-nitrooxyheptane-1 -ol, 8-nitrooxyoctane-1-ol, 9-nitrooxynonane-1 -ol and 10-nitrooxydecane-1-ol. Particularly preferred co-nitrooxyC3-ioalkane-1 -ol in the present invention is 3-nitrooxypropanol.

[0010] The term “liquid solvent” refers to solvents commonly used in feed applications, which is in liquid state at normal temperature, such as at -10-50°C, preferably at 0-30°C, more preferably at ambient temperature (i.e. at 20°C), under normal pressure such as under 1 atm. Preferred liquid solvent in the present invention is selected from the group consisting of propane-1 ,2-diol, com oil, rapeseed oil, sunflower oil, coconut oil, middle chain triglyceride (MCT) and glycerol as well as mixtures thereof. More preferred liquid solvent in the present invention is selected from the group consisting of propane-1 ,2-diol, corn oil, rapeseed oil, sunflower oil and coconut oil. Most preferred liquid solvent in the present invention is propane-1 ,2-diol and / or rapeseed oil.

[0011] The amount of the co-nitrooxyC3-ioalkane-1 -ol and the at least liquid solvent in the liquid composition of the present invention are advantageously selected such that the co-nitrooxyC3-ioalkane-1 -ol is completely dissolved in the at least liquid solvent at ambient temperature (i.e. at 20°C).

[0012] It is preferred that the amount of the co-nitrooxyC3-ioalkane-1 -ol in the liquid compositions of the present invention is at least 0.1 wt%, more preferably at least 0.5wt%, even more preferably at least 1.0wt%, such as at least 2.0wt%, at least 5.0wt%, at least 10.0wt%, at least 15.0wt%, at least 20.0wt%, at least 40.0wt%, at least 50.0wt% or even at least 60.0wt%, based on the total weight of the liquid composition. Particular advantageous ranges of the amount of the co-nitrooxyC3-ioalkane-1 -ol in the liquid composition of the present invention include 0.1 wt% to 60.0wt%, 0.5wt% to 50.0wt%, 1.0wt% to 30.0wt%, 1.5wt% to 20.0wt% and 2.0wt% to 10.0wt%, based on the total weight of the liquid composition.

[0013] It is also preferred that the total amount of the at least one liquid solvent in the liquid composition of the present invention is at least 50.0wt%, more preferably at least 60.0wt%, even more preferably at least 70.0wt%, such as at least 80.0wt%, at least 85.0wt%, at least 90.0wt%, at least 92.0wt%, at least 95.0wt%, at least 98.0wt% or even at least 99.0wt%, based on the total weight of the liquid composition. Particular advantageous ranges of the amount of the at least one liquid solvent in the liquid composition of the present invention is selected in the range of 50.0wt% to 99.9wt%, 60.0wt% to 98.0wt%, 70.0wt% to 97.0wt%, 80.0wt% to 96.0wt% and 90.0wt% to 95.0wt%, based on the total weight of the liquid composition.

[0014] In the present invention, the mass ratio of the co-nitrooxyC3-ioalkane-1 -ol to the at least one liquid solvent in the liquid composition may be selected in the range of 1 :2 to 1 :500, more preferably in the range of 1 :5 to 1 :400, most preferably in the range of 1 :10 to 1 :200, such as in the range of 1 :15 to 1 :100 (i.e. 1 mole of co-nitrooxyCs- walkane-1 -ol and 15 moles of at least one liquid solvent to 1 mole of the co-nitrooxyCs- walkane-1 -ol and 100 moles of the at least one liquid solvent).

[0015] Optionally, the liquid composition of the present application may further contain additional common used ingredients, such as formulating agents and vitamins, micro minerals, macro mineral and amino acids as well as mixtures thereof, which are suitable for a liquid formulation.

[0016] Particularly suitable formulating agents include but are not limited to water, polyols (such as glycerol and ethylene glycol), salts (such as sodium chloride, sodium benzoate and potassium sorbate), sugar and sugar derivatives (such as dextrin, glucose, sucrose and sorbitol). Preferably, the formulating agents are selected from the group consisting of water, glycerol, ethylene glycol, sodium chloride, sodium benzoate, potassium sorbate, dextrin, glucose, sucrose and sorbitol.

[0017] Particularly suitable vitamins according to the present invention are water soluble vitamins such as vitamin B12, biotin and choline, vitamin Bi , vitamin B2, vitamin Be, niacin, folic acid and panthothenate, e.g. Ca-D-panthothenate.

[0018] Particularly suitable micro minerals include but are not limited to iodine (e.g. in the form of sodium iodine), selenium, and cobalt.

[0019] Particularly suitable macro minerals include but are not limited to calcium (e.g. in the form of limestone and calcium (mono, di or triphosphate), magnesium, phosphorus and sodium (e.g. in the form of sodium chloride). In an embodiment, the liquid composition of the present invention consists essentially of:

[0020] (i) 0.1 wt% to 60wt%, preferably 0.5wt% to 50.0wt%, more preferably 1.0wt% to 30.0wt%, even more preferably 1.5wt% to 20.0wt% and the most preferably 2.0wt% to 10.0wt% of 3-nitrooxypropanol, based on the total weight of the liquid composition,

[0021] (ii) at least 50.0wt%, more preferably at least 60.0wt%, even more preferably at least 70.0wt%, such as at least 80.0wt%, at least 85.0wt%, at least 90.0wt%, at least 92.0wt%, at least 95.0wt%, at least 98.0wt% or even at least 99.0wt% of the liquid solvent, based on the total weight of the liquid composition, and

[0022] (iii) optionally, 0wt% to 3.0wt%, preferably 0wt% to 2.0wt%, most preferably 0wt% to 1 .0wt% of the additional ingredients as described herein, with the proviso that the total amount of the ingredients (i) to (iii) sum up to 100wt%.

[0023] Preferably, the liquid composition of the present invention consists essentially of the co-nitrooxyC3-ioalkane-1 -ol, and the at least one liquid solvent as described herein.

[0024] The term ‘consisting essentially of’ as used in the present invention means that the total amount of the ingredients ideally adds up to 100wt%. It is however not excluded that small amounts of impurities or additives may be present, with the proviso that the total amount of such impurities or additives is preferably less than 3wt%, more preferably less than 2wt%, most preferably less than 1wt% and which are e.g. introduced via the respective raw materials.

[0025] The liquid composition of the present invention is a clear solution, i.e. solution wherein all ingredients are fully solubilized, and which does not exhibit any undesirable opalescence, turbidity, or precipitation, and which does not contain non-solubilized materials such as silica (silicon dioxide).

[0026] The liquid composition of the present invention is also a stable solution which prevents the decomposition of 3-nitrooxypropanol, especially at elevated temperature and / or elevated concentration. Due to the enhanced stability, the liquid composition according to the present invention is particularly suitable for storing and / or transporting the co-nitrooxyCs- alkane- 1 -ol, such as preferably 3-nitrooxypropanol.

[0027] The liquid composition of the present application may be sprayed onto or mixed with animal feeds or may be added to drinking water given to animals such as ruminants to reduce formation of methane in the animals.

[0028] Thus, in the second aspect, the present invention provides the use of the liquid composition of the present invention as defined herein in animal feeds or in drinking water for reducing formation of methane in animals such as ruminants.

[0029] Furthermore, the invention provides a method of reducing the methane emission of animals such as ruminants comprising administering to the animals the liquid composition of the present invention as defined herein.

[0030] The invention is illustrated by the following Examples. All temperatures are given in °C and all parts and percentages are related to weight.

[0031] Examples

[0032] Example 1 : 3-NOP in rapeseed oil

[0033] Preparation of the liquid composition in rapeseed oil

[0034] 0.5 g, 2 g and 9 g of 3-NOP were added into refined rapeseed oil under agitation at 600 rpm at room temperature to obtain a 100 g clear solution (0.5%, 2% and 9% inclusion), respectively.

[0035] Stability study (ATR measurements)

[0036] The stability of 3-NOP in the liquid compositions as prepared above were tested by ATR measurements after 0, 5, 15 and 30 days storage at 25°C, 30°C and 40°C respectively. ln all the ATR measurements, IR spectra were measured using a Bruker Vertex 70 spectrometer equipped with an Bruker Platinum ATR accessory. Samples were measured by applying a droplet of oil on the ATR crystal. Before each measurement, a new background was collected and the sample was shaken for at least 30 seconds. The multivariate model build for quantitation purposes was based on PLS regression. The software used for model building was MATLAB Release 2021 b (The MathWorks, Inc., Natick, Massachusetts, United States). Spectral pre-treatments were done by applying a combination of 1stderivatives, standard normal variate (SNV) and mean centering (MC). A set of 120 individual samples were used for model building and validation with a split of 2 / 3 and 1 / 3, respectively.

[0037] Test results

[0038] The test results are shown in Table 1. As shown, 3-NOP were stable in the liquid composition of the present application even after 30 days storage at 25°C, 30°C and 40°C, and almost had no decomposition during the storage.

[0039] Table 1. Stability of 3-NOP in refined rapeseed oil after 0, 5, 15 and 30 days storage in different temperature conditions

[0040] Storage conditions

[0041] 3-NOP concentration Timepoints (d) 25°C 30°C 40°C

[0042] 0.5% 0 100% 100%

[0043] 5 92% 91 %

[0044] 15 93% 94%

[0045] 30 92% 92%

[0046] 2% 0 100% 100% 100%

[0047] 5 101 % 101 % 101 %

[0048] 15 100% 101 % 101 %

[0049] 30 100% 102% 101 %

[0050] 9% 0 100% 100%

[0051] 5 100% 100%

[0052] 15 100% 100%

[0053] 30 100% 100% As also shown in Figure 1 , the liquid composition of the present application containing 2% and 9% 3-NOP were still clear without any precipitation after 30 days storage at 25°C.

[0054] Example 2: 3-NOP in propane-1, 2-diol

[0055] Following the same procedures to Example 1 , 100 g clear solution containing 22% 3- NOP in propane-1 , 2-diol was prepared and the stability test was tested by ATR measurements. The results of the stability test are shown in Table 2.

[0056] Table 2. Stability of 3-NOP in propane-1 , 2-diol after 1 , 3, 6 and 9 months storage at 25°C, 60% room humidity (rh) and 30°C, 65% room humidity (rh)

[0057] Storage Conditions

[0058] Time points (month) 25°C - 60%rh 30°C - 65%rh

[0059] 3-NOP Retention in % of initial 1 103% 103%

[0060] 3 102% 103%

[0061] 6 100% 103%

[0062] 9 102% 101 %

[0063] As also shown in Figure 2, the liquid composition of the present application containingf 22% 3-NOP in propane-1 , 2-diol were still still clear without any precipitation after up to 9 months

[0064] Example 3: 3-NOP in propane-1, 2-diol in water

[0065] 3.4 g of 22% 3-NOP in propane-1 , 2-diol prepared according to Example 2 was added into 99.6 g tap water under agitation at 500 rpm at room temperature to obtain a 100 g clear solution (0.75% 3-NOP inclusion). The solution was divided in closed glass vials of 10 ml for storge. 3 repetitions per timepoint (3h, 5h, 1 d, 2d, 3d and 6d after storage) were measured on the stability of 3-NOP by using HPLC according to the below methodology.

[0066] During the test, each 10 ml glass vials containing the solution were stored open at 25°C under 60% room humidity. The concentration of 3-nitrooxypropanol was determined by HPLC using an Agilent 1260 Infinity system with an Aquasil C18, 150 x 3 mm, 3 m column and detection at 210 nm. The column oven was set to 23°C, and the autosampler had not temperature controlled. The injection volume was 10 pL. The mobile phase consisted of mobile phase A (940 mL Milli-Q-water + 60 mL acetonitrile + 1 mL methane sulfonic acid) and mobile phase B (800 mL Milli-Q-water + 200 mL acetonitrile + 1 mL methane sulfonic acid) which were used in gradient mode (0 min: 0 % B, 10 min: 0 % B, 10.5 min: 100 % B, 16 min: 100 % B, 16.5 min: 0 % B, 20 min: 0 % B (= end of run)) with a flow of 0.4 ml / min.

[0067] For calibration, solutions of 3-NOP in 80 / 20 water / acetonitrile (800 mL Milli-Q-water + 200 mL acetonitrile) at concentrations of 2, 4, 10, 20, 100 and 200 pg / mL were prepared and analysed.

[0068] The samples were prepared by weighing in duplicate about 1 g of the liquid sample into a 100 mL volumetric flask. Then, 20 mL of acetonitrile was added, followed by sonication for 10 minutes at 40°C. After cooling to room temperature, the flask was filled to volume with Milli-Q-water and vigorously mixed. An aliquot was filled into an HPLC vial prior to analysis.

[0069] The results are presented at Table 3. As shown, 3-NOP in the liquid composition of the present application is stable without any decomposition even after 3 days storage at 25°C.

[0070] Table 3. Stability of 3-NOP in propane-1 ,2-diol in tap water after 3h, 5h, 1 d, 2d, 3d and 6d storage at 25°C

[0071] Storage conditions

[0072] 3-NOP concentration Timepoints (h) 25°C

[0073] 0.75% 0 100%

[0074] 3 101%

[0075] 5 100%

[0076] 24 101%

[0077] 48 101%

[0078] 72 101%

[0079] 144 101%

[0080] Example 4: 3-NOP in other solvents According to the same procedures to Example 1 , liquid composition comprising 10% 3-NOP in other solvents (sunflower oil, corn oil, coconut oil, glycerol monosterate, palm oil fully hydrogenated) were prepared and the stability of 3-NOP was tested after 1 week storage at 3°C and 40°C. The results are shown in Table 4.

[0081] Table 4. Stability of liquid compositions containing 10% 3 NOP in different solvents after 1 week storage at 3°C and 40°C

[0082] Storage Conditions

[0083] Retention in % of initial Solvent 3°C 40°C

[0084] Sunflower oil 95% 95%

[0085] Corn oil 95% 95%

[0086] Coconut oil 79% 94%

[0087] Glycerol monosterate 24% 25%

[0088] Palm oil fully hydrogenated 67% 11% As shown, 3-NOP in the liquid compositions of the present application were stable while not in the solvents of glycerol monosterate and palm oil during storage.

Claims

Claims1. A liquid composition, which comprises a co-nitrooxyCs- alkane-l -ol dissolved in at least one liquid solvent.

2. The liquid composition of claim 1 , wherein the co-nitrooxyCs- alkane-l -ol is one or more selected from the group consisting of 3-nitrooxypropane-1 -ol, 4-nitrooxybutane- 1-ol, 5-nitrooxypentane-1-ol, 6-nitrooxyhexane-1 -ol, 7-nitrooxyheptane-1 -ol, 8-ni- trooxyoctane-1-ol, 9-nitrooxynonane-1-ol and 10-nitrooxydecane-1-ol.

3. The liquid composition of claim 1 , wherein the at least liquid solvent is one or more selected from the group consisting of propane-1 ,2-diol, com oil, rapeseed oil, sunflower oil, coconut oil, middle chain triglyceride (MCT) and glycerol.

4. The liquid composition of any of claims 1 -3, wherein the amount of the co-ni- trooxyC3- alkane-1 -ol is 0.1wt% to 60.0wt%, 0.5wt% to 50.0wt%, 1.0wt% to 30.0wt%, 1.5wt% to 20.0wt% and 2.0wt% to 10.0wt%, based on the total weight of the liquid composition.

5. The liquid composition of any of claims 1 -3, wherein the total amount of the at least one liquid solvent is at least 50.0wt%, more preferably at least 60.0wt%, even more preferably at least 70.0wt%, such as at least 80.0wt%, at least 85.0wt%, at least 90.0wt%, at least 92.0wt%, at least 95.0wt%, at least 98.0wt% or even at least 99.0wt%, based on the total weight of the liquid composition.

6. The liquid composition of any of claims 1-3, wherein the mass ratio of the co-ni- trooxyC3- alkane-1 -ol to the at least one liquid solvent is in the range of 1 :2 to 1 :500, more preferably in the range of 1 :5 to 1 :400, most preferably in the range of 1 :10 to 1 :200, such as in the range of 1 : 15 to 1 : 100.

7. The liquid composition of any of claims 1 -3, wherein the liquid composition further contains formulating agents and vitamins, micro minerals, macro mineral and amino acids as well as mixtures thereof.

8. The liquid composition of claim 7, wherein the formulating agents are selected from the group consisting of water, glycerol, ethylene glycol, sodium chloride, sodium benzoate, potassium sorbate, dextrin, glucose, sucrose and sorbitol.

9. A liquid composition, which consists essentially of:(i) 0.1 wt% to 60wt%, preferably 0.5wt% to 50.0wt%, more preferably 1.0wt% to 30.0wt%, even more preferably 1.5wt% to 20.0wt% and the most preferably 2.0wt% to 10.0wt% of 3-nitrooxypropanol, based on the total weight of the liquid composition,(ii) at least 50.0wt%, more preferably at least 60.0wt%, even more preferably at least 70.0wt%, such as at least 80.0wt%, at least 85.0wt%, at least 90.0wt%, at least 92.0wt%, at least 95.0wt%, at least 98.0wt% or even at least 99.0wt% of the liquid solvent, based on the total weight of the liquid composition, and(iii) optionally, 0wt% to 3.0wt%, preferably 0wt% to 2.0wt%, most preferably 0wt% to 1.0wt% of the additional ingredients as described herein, with the proviso that the total amount of the ingredients (i) to (iii) sum up to 100wt%.

10. Use of the liquid composition of any of claims 1 -9 in animal feeds or in drinking water for reducing formation of methane in animals such as ruminants.11 . A method of reducing the methane emission of animals such as ruminants comprising administering to the animals the liquid composition of any of claims 1-9.

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