Edible oil composition containing 7-dehydrocholesterol, previtamin D3 and vitamin D3 - Related products

Abstract

The present invention relates to an edible oil composition comprising vitamin D3, 7-dehydrocholesterol and previtamin D3, which according to the invention characteristically comprises 7-dehydrocholesterol in a mass of at least 5 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 21 mg, at least 22 mg, at least 23 mg, at least 23.5 mg, at least 24 mg, at least 24.5 mg, at least 25 mg per 100 g of the composition.

Description

[Technical field]

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

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

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

[0004] In either case, at least trace amounts of the solvent remain in the final product.

[0005] The above vitamin D3 oil compositions are not always readily absorbed by mammals, particularly humans.

[0006] They are also susceptible to oxidation of lipids, especially unsaturated lipids, by the oxygen in the air. Light also significantly degrades these oils, generating reactive species (free radicals) that sometimes initiate autocatalytic oxidation chain reactions in the oil.

[0007] Ultraviolet light (A, B and C) is known to form free radicals, which initiate a self-sustaining chemical chain reaction. When these compositions are exposed to light, they degrade very rapidly due to the rapid formation of active species (free radicals), even if they are then stored in the dark. Summary of the Invention [Problem to be solved by the invention]

[0008] One object of the present invention is to propose a new oil composition containing vitamin D3 which is more easily absorbed by the body and / or has a more stable concentration of vitamin D3 over time than oil compositions containing vitamin D3 extracted from lanolin and / or lichen. [Means for solving the problem]

[0009] The present invention relates to an edible oil composition comprising vitamin D3, 7-dehydrocholesterol and previtamin D3. Characteristically, according to the invention, it contains 7-dehydrocholesterol in an amount of 5 mg or more, 10 mg or more, 15 mg or more, 20 mg or more, 21 mg or more, 22 mg or more, 23 mg or more, 23.5 mg or more, 24 mg or more, 24.5 mg or more, 25 mg or more per 100 g of the composition.

[0010] Such oil has a stable vitamin D3 content over a long period of time. In fact, since it contains previtamin D3 and 7-dehydrocholesterol, these two compounds can be converted in situ to vitamin D3, in particular by replacing vitamin D3 degraded by oxidation. Although the applicant is not bound by the following explanation, the rearrangement reaction leading to the formation of vitamin D3 from previtamin D3 seems to be little studied, and the reaction is probably a reversible reaction leading to an equilibrium, where the reaction rate and the concentration of the species in the equilibrium depend on the medium in which the reaction occurs. Thus, the above-mentioned value of 7-dehydrocholesterol in the edible oil composition makes it possible, in particular, to easily store the composition over a long period of time and / or to promote its absorption in mammals.

[0011] The amount of 7-dehydrocholesterol per 100 g of composition may be 26 mg or less, 27 mg or less, 28 mg or less, 29 mg or less, or 30 mg or less.

[0012] The composition advantageously has a peroxide value of less than 4, in particular 2.5 or less, 2.4 or less, 2.3 or less, 2.2 or less, 2.1 or less, 2 or less, 1.9 or less, 1.8 or less, 1.7 or less, 1.6 or less, 1.5 or less, 1.4 or less, 1.3 or less, 1.2 or less, 1.15 or less or 1.10 or less, and / or 1.1 or less, 1.07 or less, 1.05 or less, 1.02 or less, 1. and anisidine values ​​of 0.00 or less, 0.97 or less, 0.95 or less, 0.90 or less, 0.87 or less, 0.85 or less, 0.82 or less, 0.80 or less, 0.77 or less, 0.75 or less, 0.72 or less, 0.70 or less, 0.67 or less, 0.65 or less, 0.62 or less, 0.60 or less, 0.57 or less, 0.55 or less, 0.52 or less, 0.50 or less, 0.47 or less, 0.45 or less, 0.43 or less, 0.40 or less, 0.37 or less, 0.35 or less, 0.33 or less, 0.30 or less, 0.27 or less, 0.25 or less, 0.22 or less, 0.20 or less, 0.17 or less, 0.15 or less, 0.12 or less, 0.10 or less, 0.07 or less, 0.05 or less, and 0.02 or less.

[0013] The above peroxide value ensures that the composition does not oxidize further as a result of reactions caused by free radicals, which are unavoidable in any case.

[0014] The anisidine value ensures minimal lipid oxidation and a stable lipid profile. The applicant has demonstrated that the lipids present are important for both the stability of Vitamin D3 and the bioavailability (absorption) of Vitamin D3 and other components that may have pharmacochemical or biological reactions.

[0015] The vitamin D3 content per 100 g of the composition according to the present invention is in all embodiments 100 μg or more, 200 μg or more, 250 μg or more, 500 μg or more, 700 μg or more, 900 μg or more, 1000 μg or more, 1200 μg or more, 1300 μg or more, 1500 μg or more, 1700 μg or more or 1900 μg or more. The vitamin D3 concentration is in all embodiments 2500 μg or less or 2000 μg or less per 100 g of the composition.

[0016] According to a first preferred embodiment, the composition according to the invention contains a vitamin D3 amount of at least 100 μg and at most 350 μg, advantageously at most 250 μg, per 100 g, and has an oxidation value of less than 1.10 and an anisidine value of less than 0.5. In this first embodiment, the amount of 7-dehydrocholesterol is comprised between 22 mg and 26 mg, and is preferably 24 mg.

[0017] According to a second preferred embodiment, the composition according to the invention comprises, per 100 g, an amount of vitamin D3 of at least 1500 μg and at most 2500 μg or at most 2000 μg, advantageously at most 1700 μg, and has an oxidation value of less than 1.15 and an anisidine value of less than 0.5. In this second embodiment, the amount of 7-dehydrocholesterol is comprised between 22 mg and 26 mg, and is preferably 23.8 mg.

[0018] According to a third preferred embodiment, the composition according to the invention comprises a vitamin D3 amount of more than 1500 μg and less than 2500 μg, advantageously 1700 μg, per 100 g, and has an oxidation value of less than 3.5, and in particular 2.5, and an anisidine value of less than 1.5, and in particular 1.1. In this third embodiment, the amount of 7-dehydrocholesterol is comprised between 22 mg and 26 mg, and is preferably 23.8 mg.

[0019] In one embodiment, which may be combined with any one of the preceding embodiments, the composition comprises at least 25% by weight polyunsaturated fatty acids and at least 40% by weight monounsaturated fatty acids, among others, these fatty acids in these ratios contributing to the bioavailability of the composition according to the invention.

[0020] Advantageously, in all embodiments of the composition according to the invention, said composition may have a cholesterol / phytosterol mass ratio greater than or equal to 135 and greater than or equal to 210, and in particular equal to 173. This ratio also contributes, in particular, to the bioavailability / absorption of vitamin D3.

[0021] 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 slow down the oxidation of the composition or contribute to the bioavailability of vitamin D3.

[0022] The composition advantageously comprises 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.

[0023] Advantageously, in all embodiments, the composition has a linoleic acid (omega 6) / alpha-linolenic acid (omega 3) mass ratio greater than or equal to 18 and less than or equal to 27.5.

[0024] Cold Extraction Advantageously, in all embodiments, the composition comprises β-sitosterol in an amount of not less than 270 μg and not more than 420 μg per 100 g of composition. β-sitosterol is known to combat hypercholesterolemia and is known to have anti-cancer and immunomodulatory properties.

[0025] Advantageously, in all of the embodiments of the emulsion, the composition comprises an oil obtained by low-temperature extraction from the larvae of beetles chosen, inter alia, from Tenebrio molitor, Alphitobius diaperinus, Tribolium castaneum and mixtures of at least two of these beetle species.

[0026] The present invention also relates to a product selected from an emulsion, a capsule, a liposome and a colloid, comprising a composition according to the invention.

[0027] definition The term "7-dehydrocholesterol" refers to a compound of formula 1:

[0028] [ka]

[0029] The term "previtamin D3" refers to a compound of formula 2:

[0030] [ka]

[0031] The previtamin D3 content can be calculated from the 7-dehydrocholesterol and vitamin D3 contents before and after irradiation. 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 disappeared as a result of irradiation and the amount of vitamin D3 produced by irradiation.

[0032] The term "vitamin D3" refers to a compound of formula 3:

[0033] [ka]

[0034] The term "edible" indicates that the composition can be ingested by a mammal, for example selected from humans, dogs, cats, horses and donkeys, for a given period of time without causing any significant problems, and indicates that the composition complies with the current regulatory requirements regarding animal and / or human nutrition, in particular the regulatory requirements regarding total oxidation values.

[0035] The term "comprising beetle oil" indicates that the composition according to the invention has the same composition as oil obtained from beetle larvae, for example by low-temperature extraction (centrifugation), except that the amount of vitamin D3 and / or 7-dehydrocholesterol and / or previtamin D3 differs from that of beetle oil. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] Two examples of producing an oil according to the invention are described below.

[0037] The larvae treatment and oil extraction steps are identical for both examples.

[0038] Only the UV treatment step is different.

[0039] Larvae treatment

[0040] Fasting process (optional)

[0041] The fasting stage for 8-12 week old larvae lasts 24 hours. This process involves draining the intestinal contents of the larvae. For this, the larvae are placed in trays (made of plastic or stainless steel). The thickness of the larvae does not exceed 1 cm. This process is carried out at temperatures between 16 and 24 °C and humidity between 35 and 70%.

[0042] Freezing process (optional)

[0043] This step allows the larvae to be stunned before the next step, i.e. killing. The step is carried out at -18°C for 5 minutes. It does not have any specific effect. The oil extraction procedure with or without this step leads to the same end product, with the same nutritional value and the same microbiological and toxicological results.

[0044] Blanching process (optional)

[0045] Fresh larvae are blanched in boiling water (100°C) for 1 min.

[0046] Extraction of the oil phase

[0047] Step 1. Centrifugation

[0048] This stage involves placing all the blanched, dehydrated or crushed larvae into a centrifuge. For one cycle, the decanter centrifuge is programmed to run for 10 minutes at a given centrifugal force. Inside the rotating conical drum of the centrifuge is a spool that rotates slower than a few rpm, pushing the solid material out of the system and allowing a solid-liquid-liquid separation: cake (solids) + oily liquid phase + aqueous liquid phase. It is the oily liquid layer that is used for the rest of the process and is called "oil".

[0049] Blanching process (optional)

[0050] After centrifugation and extraction, and passing through a filter to remove foreign particles and solid residue, the oil is collected in a container approved for contact with food.

[0051] Step 2. Heat treatment

[0052] The filtered oil obtained in this procedure is then subjected to a heat treatment by heating the oil to 90°C for a certain 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, the oil does not contain live bacteria and does not have an oxidation value (TOTOX) greater than 26.

[0053] The resulting oil contains less than 0.25 μg of vitamin D3 per 100 g of oil and approximately 24 mg of 7-dehydrocholesterol per 100 g of oil.

[0054] Vitamin D3 enrichment

[0055] Process 3.UV irradiation

[0056] The processed TM oil is first distributed in stainless steel trays (GN1 / 1, 530 x 325 x 20 mm). Stainless steel does not deteriorate due to the effects of UV light. The oil is 1 cm thick and consists of 1 kg of oil per tray. The trays containing 1 kg of TM oil are placed 21 cm under a UV neon light for 3.45 minutes for Oil 1 containing 250 μg per 100 g of oil and 30 minutes for Oil 2 containing 1700 μg per 100 g of oil in a specific room at a temperature between 20°C and 25°C and a relative humidity between 30 and 60%. The 18W UV neon light is a MIGRO UVB 310 with a spectrum of 75% UVB and 25% UVA, peak at 310 nm. The intensity of the UV light is 110 μw / cm at a distance of 21 cm. 2 reaches.

[0057] It is also possible to obtain a composition according to the invention by irradiating the oil obtained according to the method described in document WO2018122476A1.

[0058] Step 4. Centralization and homogenization

[0059] After exposure to UV light, the oil in each stainless steel tray is transferred to a covered container that is permitted for contact with food products, and the contents of the container are then agitated to ensure uniformity throughout the container.

[0060] Step 5. Filtration and packing

[0061] The oil is filtered immediately after the UV treatment and immediately packed into sealed bottles to prevent any risk of oxidation by oxygen in the air. For packing, we use disposable opaque bottles suitable for contact with food and adapted to the desired volume. Once the bottle is filled with the desired amount, the container valve is closed and the bottle is capped. Once the bottle is closed, the risk of oil contamination is practically zero. The maximum time between the end of the UV treatment and the closing of the bottle after bottling is 60 minutes.

[0062] Process 6.Storage

[0063] Before being shipped, the bottles containing the oil are stored in a special room, where the temperature is kept between 10 and 20 degrees Celsius and the humidity is kept between 35 and 70%. The room is kept dark.

[0064] Table 1 below shows the composition of the two oils, which were obtained by irradiation of Tenebrio molitor oil sold as a food product.

[0065] [Table 1]

[0066] Peroxide Value (PV) This value evaluates the degree of oxidation of unsaturated fatty acids in fats. It indicates the onset of oxidation. Peroxides are formed from free radicals that are generated in the initial phase of the oxidation reaction. The peroxide value is determined according to NF EN ISO 660 by titrimetric analysis. It is measured in meqO2 per kilogram of composition.

[0067] Anisidine value (AV) This value corresponds to the measurement of secondary oxidation products of fats. It measures the amount of aldehydes (mainly α,β-unsaturated aldehydes). The anisidine value is determined spectrophotometrically according to ISO 6885:2016.

[0068] TOTOX value (TOTal OXidation) is a measurement of the oxidation of an oil based on the peroxide value and the anisidine value. TOTOX value = (2 x PV) + AV. If the TOTOX value is above 26, the product is considered hazardous.

[0069] Sterols, including 7-DHC, a precursor of vitamin D3, are quantified by gas chromatography with flame ionization detection (GC-FID (T-AA08-WO3638)).

[0070] Lipid profiles are measured by gas chromatography with flame ionization detection (GC-FID (T-AA08-WO3638)).

[0071] Vitamin D3 is quantified by liquid chromatography with diode array detection (EN12821:2009, LC-DADLC-DAD).

[0072] Vitamin E is quantified by liquid chromatography with fluorescence detection (EN12822:2014, LC-FLD).

[0073] Fiber is determined by an enzymatic gravimetric method (COFRAC test 1-0287).

[0074] The other components are quantified by conventional methods.

[0075] Biological Data

[0076] Absorption measurements

[0077] Experimental protocol

[0078] Step 1: Animal testing

[0079] After an acclimation period, adult male Wistar rats, divided into three groups (6–8 rats per group), underwent lymphatic shunting and were then intubated with the different oils. -TM insect oil (vitamin D3 25μg / ml, referred to as "TM oil" in this report) - Lichen vitamin D3 (commercially available from D. plantes Laboratoire) diluted in fresh rapeseed oil to obtain a final vitamin D3 concentration of (25 μg / ml) (referred to in this report as "vitamin D3 lichen"). -Lanolin Vitamin D3 (sold by D. plantes Laboratoire) diluted with fresh rapeseed oil to obtain a final vitamin D3 concentration of (25 μg / ml) (referred to in this report as "Vitamin D3 Lanolin").

[0080] The oils were administered via gastric intubation to deliver the equivalent of 50 μg of vitamin D3 per rat. All three groups of mice received the same amount of vitamin D3 and oil.

[0081] A fourth control group of three saline-intubated animals also underwent lymphatic shunting, providing a reference for lymphatic fatty acid and vitamin D composition and content under fasting conditions.

[0082] 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.

[0083] Step 2: Intestinal absorption of vitamin D3

[0084] Lymph content of cholecalciferol was measured by high-performance liquid chromatography equipped with a diode array UV detector.

[0085] The intestinal absorption rate (T) of vitamin D was calculated relative to a reference value of 100% absorption: T = (RL / RH) × 100 Where: RL = amount of vitamin D in lymph / reference amount in lymph RH = amount of vitamin D in the formula taken / reference amount in the formula taken It is.

[0086] result

[0087] Lymphatic concentrations of vitamin D3 were measured for each animal group, and the results are shown in Table 2.

[0088] As expected, in the control animals, which did not receive vitamin D3, the lymph was devoid of vitamin D3.

[0089] [Table 2] Means marked with different letters are significantly different (p<0.05) by univariate ANOVA followed by Tukey's HSD test. *nd = not detected **Lichen or Lanolin Vitamin D3, diluted in fresh rapeseed oil

[0090] A better absorption of the vitamin D3 contained in the oil according to the invention can be observed.

[0091] storage

[0092] Table 3 below shows the storage results of the above oils (oil composition according to the present invention (TM), oil composition containing vitamin D3 extracted from lichen, and oil composition containing vitamin D3 extracted from lanolin).

[0093] The storage conditions are as follows: temperature: 4°C in a dark place, humidity: 50-70%

[0094] [Table 3]

[0095] Table 3 shows that the oil composition according to the invention contains the same amount of Vitamin D3 even after one month of storage under the above conditions, whereas the other two oil compositions do not.

Claims

1. An edible oil composition containing vitamin D3, 7-dehydrocholesterol, and previtamin D3, characterized in that it contains 7-dehydrocholesterol in an amount of 5 mg or more per 100 g of the composition.

2. The composition according to claim 1, characterized by having a peroxide value of less than 4 and / or an anisidine value of 1.1 or less.

3. The composition according to claim 1 or 2, characterized in that it contains 100 μg or more of vitamin D3 per 100 g of the composition.

4. The composition according to claim 1 or 2, characterized by comprising at least 25% by mass of polyunsaturated fatty acids and at least 40% by mass of monounsaturated fatty acids.

5. The composition according to claim 1 or 2, characterized by having a cholesterol / phytosterol mass ratio of 135 or more and 210 or less.

6. The composition according to claim 1 or 2, further characterized by also containing vitamin A and / or vitamin E and / or vitamin K.

7. The composition according to claim 6, 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. The composition according to claim 1 or 2, characterized by having a mass ratio of linoleic acid (omega-6) to α-linolenic acid (omega-3) of 18 or more and 27.5 or less.

9. The composition according to claim 1 or 2, characterized by containing β-sitosterol in an amount of 270 μg or more and 420 μg or less per 100 g of the composition.

10. The composition according to claim 1 or 2, characterized in that it contains an oil obtained by cold extraction from the larvae of beetles selected from Tenebrio molitor, Alphatobius diaperinus, Tribolium castaneum, and a mixture of at least two of these beetle species.

11. A product selected from emulsions, capsules, liposomes, and colloids, characterized by comprising the composition described in claim 1 or 2.