Edible oil composition containing 7-dehydrocholesterol, provitamin D3 and vitamin D3-related products

Abstract

The invention relates to an edible oil composition containing vitamin D3, 7-dehydrocholesterol and provitamin D3. Typically, according to the invention, it contains equal to or greater than 5 mg per 100 g of the composition; 10 mg; 15 mg; 20 mg; 21 mg; 22 mg; 23 mg; 23.5 mg of the raw materials; 24 mg; 24.5 mg of the raw materials; the mass of 7-dehydrocholesterol is 25 mg.

Description

Technical Field

[0001] The present invention relates to an edible oil composition containing vitamin D3. Prior art

[0002] Vitamin D3 is commonly extracted from lanolin or lichen.

[0003] Extraction of vitamin D3 from lanolin or lichen requires the use of a solvent. The vitamin D3 extracted from lanolin or lichen is then mixed with a vegetable oil, typically rapeseed oil.

[0004] In both cases, the solvent remains in the finished product at least in trace amounts.

[0005] The above-mentioned vitamin D3 oil compositions are not always easily absorbed by mammals, especially humans.

[0006] Since lipids (particularly unsaturated lipids) are oxidized by oxygen in the air, they are also vulnerable. Light also causes these oils to deteriorate significantly, generating activated species (free radicals) that produce oxidation chain reactions that are sometimes self-catalyzed in the oil.

[0007] It is known that ultraviolet rays (A, B and C) form free radicals which cause self-sustaining chemical chain reactions. When these compositions are exposed to light, they deteriorate very quickly due to the rapid formation of activated species (free radicals), even if they are subsequently stored in the dark.

[0008] An object of the present invention is to provide a novel oil composition containing vitamin D3 which is easily absorbed by the body and / or which has a more stable vitamin D3 concentration over time than an oil composition containing vitamin D3 extracted from lanolin and / or lichen. SUMMARY OF THE INVENTION

[0010] The present invention relates to an edible oil composition containing vitamin D3, 7-dehydrocholesterol and provitamin D3. Typically, according to the present invention, it contains 7-dehydrocholesterol in an amount equal to or greater than 5 mg; 10 mg; 15 mg; 20 mg; 21 mg; 22 mg; 23 mg; 23.5 mg; 24 mg; 24.5 mg; 25 mg per 100 g of the composition.

[0011] Such oils have a vitamin D3 content that is stable over time. In fact, since they contain provitamin D3 and 7-dehydrocholesterol, these two compounds can be converted into vitamin D3 in situ, replacing the vitamin D3 that has deteriorated, particularly by oxidation. The applicant is not bound by the following explanation, and the rearrangement reaction that leads to the formation of vitamin D3 from provitamin D3 does not seem to have been studied in large quantities; it may be a reversible reaction that leads to equilibrium, wherein the kinetics and concentration of the species at equilibrium depend on the medium in which it occurs. Thus, the above-mentioned value of 7-dehydrocholesterol in the edible oil composition enables the composition to be easily preserved over time and / or helps it to be absorbed, particularly in mammals.

[0012] The amount of 7-dehydrocholesterol per 100 g of the composition may be less than or equal to 26 mg, 27 mg, 28 mg, 29 mg or 30 mg.

[0013] The composition advantageously has a peroxide value of less than 4, in particular less than or equal to 2.5; 2.4; 2.3; 2.2; 2.1; 2; 1.9; 1.8; 1.7; 1.6; 1.5; 1.4; 1.3; 1.2; 1.15 or 1.10 and / or less than or equal to 1.1; 1.07; 1.05; 1.02; 1.00; 0.97; 0.95; 0.90; 0.87; 0.85; 0.82; 0.80; 0. The anisidine values ​​are 77; 0.75; 0.72; 0.70; 0.67; 0.65; 0.62; 0.60; 0.57; 0.55; 0.52; 0.50; 0.47; 0.45; 0.43; 0.40; 0.37; 0.35; 0.33; 0.30; 0.27; 0.25; 0.22; 0.20; 0.17; 0.15; 0.12; 0.10; 0.07; 0.05; 0.02.

[0014] The above mentioned peroxide values ​​ensure that the composition will not be further oxidized by reactions caused by free radicals, which are unavoidable in any case.

[0015] The anisidine value ensures minimal lipid oxidation and a stable lipid profile (lipid composition). Applicants have demonstrated that the lipids present are important for both the stability of Vitamin D3 and for the bioavailability (absorption) of Vitamin D3 and any other components that may have a pharmaceutical or biological response.

[0016] In all embodiments, the content of vitamin D3 in the composition according to the present invention per 100 g is greater than or equal to 100 μg, 200 μg, 250 μg, 500 μg, 700 μg, 900 μg, 1000 μg, 1200 μg, 1300 μg, 1500 μg, 1700 μg or 1900 μg. In all embodiments, for 100 g of the composition, the vitamin D3 concentration is less than or equal to 2500 μg or 2000 μg.

[0017] According to a first preferred embodiment, the composition according to the invention contains vitamin D3 in an amount greater than or equal to 100 μg and less than or equal to 350 μg and advantageously equal to 250 μg per 100 g, with an oxidation value less than 1.10 and an anisidine value less than 0.5. In this first embodiment, the amount of 7-dehydrocholesterol is between 22 mg and 26 mg and is preferably equal to 24 mg.

[0018] According to a second preferred embodiment, the composition according to the invention contains vitamin D3 in an amount greater than or equal to 1500 μg and less than or equal to 2500 μg or 2000 μg and advantageously equal to 1700 μg per 100 g, with an oxidation value less than 1.15 and an anisidine value less than 0.5. In this second embodiment, the amount of 7-dehydrocholesterol is between 22 mg and 26 mg and is preferably equal to 23.8 mg.

[0019] According to a third preferred embodiment, the composition according to the invention contains vitamin D3 in an amount greater than or equal to 1500 μg and less than or equal to 2500 μg and advantageously equal to 1700 μg per 100 g, and has an oxidation value less than or equal to 3.5 and in particular equal to 2.5 and an anisidine value less than or equal to 1.5 and in particular equal to 1.1. In this third embodiment, the amount of 7-dehydrocholesterol is between 22 mg and 26 mg and is preferably equal to 23.8 mg.

[0020] In one embodiment that may be combined with any of the preceding embodiments, the composition contains at least 25% by mass of polyunsaturated fatty acids and at least 40% by mass of monounsaturated fatty acids. In particular, the inclusion of these fatty acids in these proportions contributes to the bioavailability of the composition according to the invention.

[0021] Advantageously, in all embodiments of the composition according to the invention, it may have a cholesterol / phytosterol mass ratio equal to or greater than 135 and equal to or less than 210, and in particular equal to 173. This ratio also contributes particularly to the bioavailability / absorption of vitamin D3.

[0022] Advantageously, in all embodiments, the composition also contains vitamin A and / or vitamin E and / or vitamin K. Vitamins A and K are antioxidants that will help slow oxidation of the composition or aid in the bioavailability of the vitamin D3.

[0023] It advantageously contains at least 15 μg of vitamin E per 100 g of composition, and in particular at least 3.00 μg of vitamin K1 per 100 g of composition.

[0024] Advantageously, in all embodiments, it has a linoleic acid (ω6) / α-linolenic acid (ω3) mass ratio equal to or greater than 18 and equal to or less than 27.5.

[0025] Advantageously, in all embodiments, it contains beta-sitosterol in an amount greater than or equal to 270 μg and less than or equal to 420 μg per 100 g of composition. Beta-sitosterol is known to combat hypercholesterolemia; it is known to have anticancer and immunomodulatory properties.

[0026] Advantageously, in all embodiments, it contains oil obtained by cold extraction from beetle larvae, in particular chosen from Tenebrio molitor, Alphitobius diaperinus, Tribolium castaneum and a mixture of at least two of these beetle species.

[0027] The invention also relates to a product selected from emulsions, capsules, liposomes and colloids, containing the composition according to the invention.

[0028] definition

[0029] The term 7-dehydrocholesterol constitutes the compound of formula 1 below:

[0030] [Formula 1]

[0031]

[0032] The term previtamin D3 refers to a compound of the following formula 2:

[0033] [Formula 2]

[0034]

[0035] The previtamin D3 content can be calculated from the contents of 7-dehydrocholesterol and vitamin D3 before and after the irradiation treatment. The maximum amount of vitamin D3 that can be produced by irradiation is equal to the amount of 7-dehydrocholesterol contained before irradiation. The amount of previtamin D3 corresponds to the difference between the amount of 7-dehydrocholesterol that disappears due to irradiation and the amount of vitamin D3 produced by irradiation.

[0036] The term vitamin D3 refers to the compound of the following formula 3:

[0037] [Formula 3]

[0038]

[0039] The term "edible" means that the composition can be ingested within a given period by a mammal chosen for example from humans, dogs, cats, horses and donkeys without causing any significant problems; the composition complies with the current regulatory conditions for use in animal and / or human nutrition, in particular as regards the gross oxidative value.

[0040] The expression "containing beetle oil" means that the composition according to the invention has the same composition as the oil obtained, for example, by cold extraction (centrifugation) from beetle larvae, except for the amount of vitamin D3 and / or 7-dehydrocholesterol and / or previtamin D3, which is different from the amount of beetle oil.

[0041] Detailed description of the invention - Examples

[0042] Two examples of producing oils according to the invention will now be described.

[0043] Larval handling and oil extraction stages were identical for both examples.

[0044] Only the UV processing stage is different.

[0045] Larvae treatment

[0046] Fasting Steps (Optional)

[0047] The fasting phase for larvae between 8 and 12 weeks old lasts 24 hours. This step involves emptying the intestinal contents of the larvae. To do this, the larvae are placed in trays (plastic or stainless steel). The thickness of the larvae does not exceed 1 cm. It is carried out at a temperature of 16 to 24°C and a humidity of 35 to 70%.

[0048] Freezing step (optional)

[0049] This step allows the larvae to be stunned before the following step: slaughter. This step is carried out at -18°C for 5 minutes. This step does not have any specific effects. The oil extraction procedure with or without this step produces the same end product, with the same nutritional value and the same microbiological and toxicological results.

[0050] Blanching step (optional)

[0051] Fresh larvae were blanched in boiling water (100°C) for one minute.

[0052] Oil extraction stage

[0053] Step 1. Centrifugation

[0054] This stage involves placing whole, blanched, dehydrated or crushed larvae in a centrifuge. For one cycle, the decanter centrifuge is programmed to run for 10 minutes at a given centrifugal force. Inside the rotating conical bowl of the centrifuge is a spool that rotates at a speed of a few rpm, pushing solid material out of the system and allowing solid-liquid-liquid separation: cake (solids) + oily liquid phase + aqueous liquid phase. The oily liquid phase is used in the rest of the process and is called "oil".

[0055] Blanching step (optional)

[0056] After centrifugation and extraction, the oil is collected in containers allowed to come into contact with food, passing through filters to remove foreign matter and solid residues.

[0057] Step 2. Heat treatment

[0058] The filtered oil obtained in this way is subsequently subjected to a heat treatment which heats the oil to 90°C for a period of time to ensure that the oil complies with regulations; at this stage, the oil is considered a processed product suitable for human or animal consumption. In particular, it does not contain live bacteria and does not have an oxidation value (TOTOX) higher than 26.

[0059] The oil obtained contained less than 0.25 μg of vitamin D3 / 100 g of oil and approximately 24 mg of 7-dehydrocholesterol / 100 g of oil.

[0060] Vitamin D3 Enriched

[0061] Step 3. UV exposure

[0062] The treated TM oil was first distributed in stainless steel trays (GN 1 / 1; 530×325×20 mm). Stainless steel does not deteriorate under the action of UV light. The oil was 1 cm thick and each tray contained 1 kg of oil. The trays containing 1 kg of TM oil were then placed under UV neon light at 21 cm, at a temperature of 20°C to 25°C and a relative humidity of 30 to 60% in a specific room for 3.45 minutes for Oil 1 containing 250 μg / 100 g of oil and 30 minutes for Oil 2 containing 1700 μg / 100 g of oil. 18W UV neon lamp (MIGRO UVB 310 with 75% UVB and 25% UVA, peak spectrum at 310 nm). The UV light intensity reached 110 μW / cm at 21 cm. 2 .

[0063] The composition according to the invention can also be obtained by irradiating the oil obtained according to the method described in document WO 2018122476 A1.

[0064] Step 4. Concentration and homogenization

[0065] After UV exposure, the oil in each stainless steel tray was transferred to a container with a lid that allowed contact with food. The contents of the container were then stirred to achieve uniformity throughout the container.

[0066] Step 5. Filtering and packaging

[0067] The oil is filtered immediately after the UV treatment and immediately packaged in sealed bottles to prevent any risk of oxidation by oxygen in the air. For packaging, we use disposable opaque bottles suitable for contact with food and adapted to the required volume. Once the bottle is filled with the required volume, the reservoir valve is closed and the bottle is capped. Once the bottle is closed, the risk of oil contamination is almost zero. The maximum time between the end of the UV treatment and the closure of the bottle after bottling is 60 minutes.

[0068] Step 6. Storage

[0069] Before shipment, the oil bottles are stored in a dedicated room. The ambient temperature is 10 to 20°C and the humidity is 35 to 70%. The room is kept dark.

[0070] The composition of the two oils is shown in Table 1 below. The two oils were obtained by irradiating mealworm oil sold as a food product.

[0071] [Table 1]

[0072]

[0073] Peroxide value (PV). This value evaluates the degree of oxidation of unsaturated fatty acids in fats. This value indicates the onset of oxidation. Peroxides are formed from free radicals generated in the initiation phase of the oxidation reaction. The peroxide value is determined by titration according to NF EN ISO 660. It is measured in meqO2 / kg of composition.

[0074] Anisidine value (AV). This value corresponds to the measured value of secondary oxidation products of fats. This value measures the amount of aldehydes (mainly α,β-unsaturated aldehydes). The Anisidine value is measured spectrophotometrically according to ISO 6885:2016.

[0075] The TOTOX value (Total Oxidation) is a measure of the oxidation of the oil based on the Peroxide Value and the Anisidine Value. TOTOX value = (2 x PV) + AV. If the TOTOX value is greater than 26, the product is considered hazardous.

[0076] Sterols including 7-DHC, a precursor of vitamin D3, were quantified by gas chromatography with a flame ionization detector (GC-FID (T-AA08-WO3638)).

[0077] Lipid profiles were determined by gas chromatography with a flame ionization detector (GC-FID (T-AA08-WO3638)).

[0078] Vitamin D3 was quantified by liquid chromatography with diode array detection (EN 12821:2009, LC-DADLC-DAD).

[0079] Vitamin E was quantified by liquid chromatography with fluorescence detection (EN 12822:2014, LC-FLD).

[0080] Fiber was quantified by enzymatic gravimetric method (COFRAC test 1-0287).

[0081] Other components were quantified by conventional methods.

[0082] Biological data

[0083] Absorption measurement

[0084] Experimental protocol

[0085] Step 1: Animal Experiment

[0086] After an acclimation period, three groups of adult male Wistar rats were subjected to lymphatic shunt surgery (6 to 8 rats per group) and then cannulated with different oils.

[0087] -TM insect oil (25 μg / ml vitamin D3, referred to as "TM oil" in this report)

[0088] - Lichen Vitamin D3 (sold by D. plants Laboratoire), diluted in virgin rapeseed oil to obtain a final vitamin D3 concentration of (25 μg / ml) (referred to as "VitD3 Lichen" in this report)

[0089] - Lanolin Vitamin D3 (sold by D. plants Laboratoire), diluted in virgin rapeseed oil to obtain a final Vitamin D3 concentration of (25 μg / ml) (referred to as "VitD3 Lanolin" in this report)

[0090] Each oil was administered via gastric cannula to provide the equivalent of 50 μg vitamin D3 / rat. All 3 groups of mice received the same amount of vitamin D3 and oil.

[0091] A fourth control group of 3 animals cannulated with saline also underwent lymphatic shunting. This group will serve as a reference for the fatty acid and vitamin D composition and content of lymph under fasting conditions.

[0092] For each group of animals, lymph fluid was collected over a 6-hour period to determine its vitamin D3 concentration as a function of the oil ingested.

[0093] Step 2: Intestinal absorption of vitamin D3

[0094] Lymphatic content of cholecalciferol was determined by high performance liquid chromatography equipped with a diode array UV detector.

[0095] The intestinal absorption rate (T) of vitamin D was calculated by comparison with a reference substance having an absorption rate of 100%:

[0096] T=(RL / RH)×100

[0097] in:

[0098] RL = amount of vitamin D in lymph / reference amount in lymph

[0099] RH = Amount of vitamin D in the preparation taken / reference amount in the preparation taken

[0100] result

[0101] The lymphatic concentration of vitamin D3 was measured in each group of animals. The results are shown in Table 2.

[0102] As expected, in control animals that were not given vitamin D3, the lymph was devoid of vitamin D3.

[0103] [Table 2]

[0104]

[0105] Means marked with different letters are significantly different (p < 0.05), univariate analysis of variance followed by Tukey HSD test

[0106] *nd=Not Detectable

[0107] **Lichen or lanolin vitamin D3, diluted in virgin canola oil

[0108] It can be observed that the absorption of the vitamin D3 contained in the oil according to the invention is better.

[0109] store

[0110] Table 3 below shows the storage results of the above oils (the oil composition according to the present invention (TM), the oil composition containing vitamin D3 extracted from lichen, and the oil composition containing vitamin D3 extracted from lanolin).

[0111] Storage conditions are as follows: Temperature: 4°C in the dark, Humidity: 50 to 70%

[0112] [Table 3]

[0113] D3 T=0(%T0) D3 T=1M(%T0) D3 TM 100 100 D3 Lichen 100 72.72727273 D3 Lanolin 100 85.95744681

[0114] Table 3 shows that the oil composition according to the present invention still contains the same amount of vitamin D3 even after storage for one month under the above conditions, which is not the case for the other two oil compositions.

Claims

1. An edible oil composition containing vitamin D3, 7-dehydrocholesterol and provitamin D3, characterized in that It contains 7-dehydrocholesterol in a mass equal to or greater than 5 mg; 10 mg; 15 mg; 20 mg; 21 mg; 22 mg; 23 mg; 23.5 mg; 24 mg; 24.5 mg; 25 mg per 100 g of composition.

2. The composition according to claim 1, characterized in that It has a peroxide value of less than 4, in particular less than or equal to 2.5; 2.4; 2.3; 2.2; 2.1; 2; 1.9; 1.8; 1.7; 1.6; 1.5; 1.4; 1.3; 1.2; 1.15 or 1.10 and / or less than or equal to 1.1; 1.07; 1.05; 1.02; 1.00; 0.97; 0.95; 0.90; 0.87; 0.85; 0.82; 0.80; 0.77; 0.75; 0.72; 0.70; 0.67; 0.65; 0.62; Anisidine values ​​of 0.60; 0.57; 0.55; 0.52; 0.50; 0.47; 0.45; 0.43; 0.40; 0.37; 0.35; 0.33; 0.30; 0.27; 0.25; 0.22; 0.20; 0.17; 0.15; 0.12; 0.10; 0.07; 0.05; 0.

02.

3. The composition according to claim 1 or 2, characterized in that It contains vitamin D3 in an amount equal to or greater than 100 μg, 200 μg, 250 μg, 500 μg, 700 μg, 900 μg, 1000 μg, 1200 μg, 1300 μg, 1500 μg, 1700 μg or 1900 μg per 100 g of the composition.

4. The composition according to any one of the preceding claims, characterized in that It contains at least 25% by mass of polyunsaturated fatty acids and at least 40% by mass of monounsaturated fatty acids.

5. A composition according to any one of the preceding claims, characterised in that It has a cholesterol / phytosterol mass ratio equal to or greater than 135 and equal to or less than 210, and in particular equal to 173.

6. A composition according to any one of the preceding claims, characterised in that It also contains vitamin A and / or vitamin E and / or vitamin K.

7. The composition according to claim 7, characterized in that It contains at least 15 μg of vitamin E per 100 g of the composition, and in particular at least 3.00 μg of vitamin K1 per 100 g of the composition.

8. A composition according to any one of the preceding claims, characterised in that It has a linoleic acid (ω6) / α-linolenic acid (ω3) mass ratio of equal to or greater than 18 and equal to or less than 27.

5.

9. A composition according to any one of the preceding claims, characterised in that It contains β-sitosterol in an amount greater than or equal to 270 μg and less than or equal to 420 μg per 100 g of the composition.

10. A composition according to any one of the preceding claims, characterised in that It contains oil obtained by cold extraction from beetle larvae, in particular chosen from Tenebrio molitor, Alphitobius diaperinus, Tribolium castaneum and mixtures of at least two of these beetle species.

11. A product selected from the group consisting of emulsions, capsules, liposomes and colloids, characterized in that It contains a composition according to any one of the preceding claims.

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