JP7504609B2 - Composition containing lignan compounds dissolved in clove oil, and method for increasing solubility of sesamin - Google Patents

Description

Lignan compounds contained in sesame, such as sesamin and sesamolin, have been reported to have cholesterol-lowering effects (Non-Patent Document 1), antihypertensive effects (Non-Patent Documents 2 and 3), antioxidant effects (Non-Patent Document 4), liver protective effects (Non-Patent Document 5), liver cancer prevention effects (Non-Patent Document 6), breast cancer prevention effects (Non-Patent Document 7), and immunostimulatory effects (Non-Patent Document 8).
Sesamin is fat-soluble and is said to dissolve in oils and fats, but in reality, it only dissolves about 1% in common edible oils. Therefore, commercially available sesamin preparations either dissolve sesamin in edible oils by reducing the sesamin content, or disperse or suspend sesamin. In the former case, the content of the ingredient per capsule is low, and in the latter case, the capsule content liquid becomes opaque, making it less attractive in appearance and less appealing to consumers. An example of the former is Suntory's "Sesamin EX," a soft capsule with a transparent content liquid. This soft capsule uses rice germ oil to dissolve 3.3 mg of sesamin per 370 mg capsule, and the content ratio of sesamin in the capsule content liquid is 0.89%. In addition, Kadoya Oil Co., Ltd.'s "Sesame Sesamin" is also a soft capsule with a transparent content liquid, and sesame oil is used to dissolve 5 mg of sesamin per 200 mg of the content liquid per capsule, with 2.5% of sesamin dissolved in the capsule content liquid. An example of the latter is "Black Sesame Sesamin Supplement 90 Capsules" by Nichie Co., Ltd. This is a soft capsule with a white membrane, and 15 mg of sesamin is suspended in sesame oil using vegetable lecithin. Also, "Sesamin Capsule" by Organland Co., Ltd. is a soft capsule with a yellow membrane, and 10 mg of sesamin is suspended in sesame oil using emulsifier and beeswax.
MCT is widely known as an edible fat capable of dissolving sesamin (Patent Document 1). In addition, there are examples of blends of γ-oryzanol and rice germ oil (Patent Document 2), epigallocatechin gallate (Patent Document 3), and quercetin (Patent Document 4) with the aim of increasing the degree of absorption in the body by combining with sesamin. However, even with these blends, it is difficult to increase the sesamin concentration while maintaining the transparency of the capsule content liquid, and the concentration at which the capsule content liquid containing sesamin can be kept transparent is about 1 to 2.5% in the capsule content liquid.
As a result, the ingredient content per capsule is low, which results in an increase in the number of capsules to be orally administered, resulting in higher costs. At the same time, the number of capsules to be ingested is too high, imposing a burden on users when taking the medicine.

Patent No. 5666760 Patent No. 6157463 Patent No. 5547891 Patent No. 5276316

Lipid Res. , 1991 Apr;32(4):629-38. Biol. Pharm. Bull. , 1995 Jul;18(7):1016-9. Biol. Pharm. Bull. , 1995 Sep;18(9):1283-5. Biofactors, 2004;21(1-4):191-6. Ann. Nutr. Metab. , 1993;37:218-24. Proc. Am. Assoc. Cancer Res. , 2000;41:459. Anticancer Res. , 1992;12:1259. Biosci. Biotechnol. Biochem., 1994;59:1855-58.

As mentioned above, it is not possible to dissolve lignan compounds such as sesamin and sesamolin in high concentrations while maintaining the transparency of the capsule content liquid, so it has been necessary to either disperse or suspend sesamin powder to produce a composition that contains a large amount of sesamin and is opaque in appearance, or to reduce the sesamin content per capsule to produce a transparent composition in which sesamin is dissolved. In other words, there is a demand for a composition or capsule that maintains transparency while dissolving sesamin in a high concentration per capsule in order to reduce the number of capsules that need to be ingested while still being transparent.

After screening various solvents, we found clove oil as a solvent that can dissolve lignan compounds such as sesamin and sesamolin at high concentrations without causing crystallization, and thus completed the present invention.
That is, the present invention is as follows.
(1) A composition containing lignan compounds including sesamin and sesamolin dissolved in clove oil.
(2) The composition described in (1), wherein the concentration of sesamin dissolved in the composition is 2% by mass to 25% by mass.
(3) A food or beverage containing the composition described in (1) or (2).
(4) The food or beverage described in (3) above, which is in the form of a capsule.
(5) A method of increasing the solubility of sesamin in clove oil using sesamolin.

The composition of the present invention increases the amount of ingredients ingested per capsule while maintaining the transparency of the capsule contents compared to conventional formulations formulated by dispersion and suspension or formulations dissolved in edible oil, thereby reducing the number of capsules to be orally ingested. This also allows for a reduction in capsule processing costs.

<Lignan compounds>
In the present invention, the lignan compound is a type of phenylpropanoid, a polyphenol compound derived from a plant-derived dibenzylbutane skeleton. Lignan compounds are extracted from red sesame (linseed), sesame, and Brassica vegetables, and in some cases, purified ones can be used. Examples include sesaminol, pinoresinol, syringaresinol, sesamin, sesamolin, lariciresinol, secoisolariciresinol, matairesinol, and hydroxymatairesinol.

As mentioned above, lignan compounds such as sesamin and sesamolin contained in sesame have been reported to have cholesterol-lowering, antihypertensive, antioxidant, liver-protecting, liver cancer-preventing, breast cancer-suppressing, and immunostimulating effects. Many soft capsules containing sesamin currently on the market set the daily sesamin intake at 10-15 mg. In fact, it has been reported that when people who felt strong fatigue took soft capsules containing 10 mg of sesamin and vitamin E for eight consecutive weeks, significant improvements were observed in fatigue, beauty, and sleep, and the LDL oxidation lag time was increased, leading to increased antioxidant activity in the blood. (Glob. J. Health Sci., 2015; 7: 1-10).

<Sesamin>
Sesamin is a lignan compound found in large amounts in sesame oil and Japanese pepper, and has the following structural formula.

<Sesamolin>
Like sesamin, sesamolin is a lignan compound found in large amounts in sesame oil and Japanese pepper, and has the following structural formula.

<Sesame extract>
When sesamin or sesamolin is used as the lignan compound in the lignan compound-containing composition of the present invention (hereinafter, also referred to simply as the "composition of the present invention." In addition, "a composition containing a lignan compound dissolved in clove oil,""a capsule content liquid composition," and "a supernatant composition" are also included in the "lignan compound-containing composition of the present invention"), it is preferable to use a sesamin concentrate obtained by extraction and/or purification from a food-derived material such as sesame oil, for example, a sesame extract.
The concentrations of sesamin and sesamolin in sesame seeds have been reported (Bull. Natl. Inst. Crop Sci., 2008; 9:27-61). According to this document, the sesamin:sesamolin concentration ratio in sesame seeds varies depending on the variety. For example, the sesamin and sesamolin concentrations in sesame extract Sesamin 90 (Coach Boueki) are 82.3% sesamin and 1.6% sesamolin, SP-080A (Arjuna Natural Extracts, Aruba, India) are 67.5% sesamin and 18.4% sesamolin, and SP-811 (Arjuna Natural Extracts, Aruba, India) are 39.7% sesamin and 46.5% sesamolin.

<Clove oil>
In the present invention, the lignan compounds are used by dissolving them in clove oil.

The clove oil used in the composition of the present invention is not limited in any way by its origin, form, production method, or the like.
For example, clove oil is obtained by steam distillation of the leaves and buds of Syzygium aromaticum, but also includes oils obtained from different parts of the same plant and from related species of the Myrtaceae genus.

<Lignan compound-containing composition>
The composition of the present invention is a composition containing lignan compounds dissolved in clove oil, and the lignan compounds are preferably sesamin and sesamolin.
The composition of the present invention contains sesamin in an amount of preferably 2 to 25% by mass, more preferably 3 to 20% by mass, based on the total mass of the composition. Conventionally, the sesamin concentration at which a composition containing dissolved sesamin can be kept transparent was about 1 to 2.5% by mass in the composition, but by selecting clove oil as a solvent for dissolving lignan compounds, the sesamin concentration in the composition can be increased.

<Method for preparing a lignan compound-containing composition>
The sesame extract is mixed with clove oil and dissolved by heating, preferably at 75 to 100° C. for 5 to 15 minutes, more preferably at 85° C. for 10 minutes. Thereafter, the solution is cooled to room temperature with water and allowed to stand overnight. After standing, the sample is centrifuged at 15° C. and 10,000 rpm for 5 minutes, and the resulting supernatant is the lignan compound-containing composition of the present invention.
The concentrations (mass %) of sesamin and sesamolin contained in this lignan compound-containing composition are quantified by liquid chromatography (LC).

<Lignan compound-containing composition and food and drink>
The composition of the present invention is preferably provided in the form of a food or drink (functional food, dietary supplement, nutrient functional food, special purpose food, food for specified health uses, nutritional supplement, dietary food, health food, supplement, etc.) and a pharmaceutical. When the composition of the present invention is provided as a food or drink, the form may be provided in the form of a health food such as a tablet, capsule, powder, granule, drink (including a solution and a suspension), or in the form of a soft drink, tea drink, dairy product such as yogurt or lactic acid bacteria drink, seasoning, processed food, dessert, confectionery (e.g., gum, candy, jelly), etc., but is not limited thereto.

In addition to sesamin and clove oil, the composition of the present invention may contain any other desired ingredients, so long as the effects of the composition are not impaired. For example, vitamins such as vitamin D and vitamin E, fats and oils such as DHA and EPA, minerals, hormones, nutritional ingredients, fragrances, and other physiologically active ingredients may be added, as well as preservatives, stabilizers, antioxidants, and the like that are added during formulation.

<Transparency assessment>
The composition of the present invention can dissolve lignan compounds (especially sesame extract) at high concentration and prepare a transparent solution. Transparency can be evaluated by total light transmission method, cloudiness (haze) method, transparency method using a see-through clock, or other turbidity method, or by a method of evaluating transmittance using a spectrophotometer. The composition of the present invention has a feature that the transparency measured by the above method while maintaining a high sesamin concentration is comparable to that of existing capsule contents with low sesamin concentration, such as Sesamin EX and Sesamin from Kobayashi Pharmaceutical. In addition, when a composition containing lignan compounds is filled into a soft capsule with a transparent coating, transparency is important because the transparent content liquid has the advantage of making the appearance beautiful and improving consumer appeal.
When the transparency is evaluated by the transmittance at 750 nm using an ultraviolet-visible spectrophotometer, the transparency of the composition of the present invention is preferably a transmittance at 750 nm of 80% or more, particularly preferably 95% or more.

The following provides further explanation with reference to examples, but the present invention is not limited to these examples.

(Solubility test)
1. Preparation of Experimental Materials Sesame extracts used were Sesamin 90 purchased from Coach Boueki, and SP-080A or SP-811 procured from Arjuna Natural Extracts (Aluva, India). The sesamin and sesamolin concentrations in each product are shown in Table 1.

Clove oil was purchased from PT Indesso Aroma and was obtained by steam distillation of Eugenia caryophyllata leaves.
MCT was purchased from Eiwa Trading Co., Ltd. (Tokyo, Japan) and was prepared by squeezing the fruits of Elaeis guineensis.
Rice germ oil was purchased from Tsuno Life Sciences Co., Ltd. and used.
The sesame oil used was purchased from Takemoto Oil Co., Ltd. under the name "Taihaku Sesame Oil."

2. Evaluation method for solubility test The solubility of sesamin in the supernatant after centrifugation was evaluated using the following four-level scale (◎, ◯, △, ×).

◎: Sesamin concentration is 10% by mass or more. ◯: Sesamin concentration is 3% by mass or more and less than 10% by mass. △: Sesamin concentration is 1% by mass or more and less than 3% by mass. ×: Sesamin concentration is less than 1% by mass.

(LC-MS analysis method)
The concentrations of sesamin and sesamolin in the samples were quantified using a UPLC-MS system (LCMS-8050, Shimadzu Corporation).
The quantitative analysis of sesamin was carried out under the following conditions. The column used was a Shim-pack XR-ODSII (2.0 mm ID × 75 mm, particle size; 2.2 μm, Shimadzu Corporation). The flow rate was 0.2 mL/min. The mobile phase was a 4:1 mixture of methanol and 5 mM ammonium acetate aqueous solution. The retention time was 2.25 min. The injected solution volume was 1 μL. The ESI-MS measurement was carried out under the following conditions. The drift voltage was 2.5 kV. The ionization mode was positive mode. The ion source temperature was 100°C. The desolvation gas flow rate was 10 L/min, and the desolvation temperature was 100°C. The collision-induced dissociation gas used was argon gas at 270 kPa, and the collision energy was 11 eV. The compound was identified by multiple reaction monitoring, which tracked the transition from [M+NH ₄ ] ⁺ =m/z 372.20 to [M+NH ₄ --C ₇ H ₅ O ₂ --H ₂ O] ⁺ =m/z 372.20.
Sesamolin was quantified using the same method as the above sesamin quantification method, except for the following points. The mobile phase was methanol:5 mM ammonium acetate aqueous solution, linearly changed in concentration from 4:1 to 1:0 in 2.5 minutes, and then back to 4:1 in 2.51 minutes. The retention time was 2.55 minutes. ESI was introduced using MS/MS. The drift voltage was 4.0 kV. The ion source temperature was 225°C, and the desolvation temperature was 150°C. The collision energy of argon gas was 27 _eV . Multiple reaction monitoring was used to identify the compound, _{and the transition from [M+NH4]+ = m/z 388.20 to [M-C12H12O5 +} ^H _] ⁺ = m/z 135.00 was tracked.

(Example 1)
Any one of the sesame extracts A to C shown in Table 1 was mixed with essential oil (clove oil) in the composition ratio shown in Table 2, and dissolved by heating at 85° C. for 10 minutes. The compositions (theoretical values) of sesamin and sesamolin contained in the blended composition at the time of blending were calculated based on Table 1 and are shown in Table 2.
The resulting solutions were then cooled to room temperature with water and allowed to stand overnight. Since some samples showed precipitation during standing, the solutions were centrifuged at 15°C and 10,000 rpm for 5 minutes using a multipurpose centrifuge CAX-371 (Tomy Seiko Co., Ltd.), and the concentrations (mass%) of sesamin and sesamolin in the supernatant were quantified using a UPLC-MS system (LCMS-8050, Shimadzu Corporation). The results are shown in Table 3.

The solubility of sesamin could be increased by using clove oil as a solvent, regardless of the sesamin:sesamolin concentration ratio, and sesamin could be dissolved in the supernatant composition up to a maximum of 19.0% by mass.

(Reference Example 1)
The same procedure as in Example 1 was carried out, except that the sesame extract and MCT were mixed in the composition ratio shown in Table 4. MCT was purchased from Eiwa Trading Co., Ltd. (Tokyo, Japan).
The compositions (theoretical values) of sesamin and sesamolin contained in the blended composition at the time of blending are shown in Table 4, and the concentrations of sesamin and sesamolin contained in the supernatant (measured by LC) are shown in Table 5.

By setting the sesamin:sesamolin concentration ratio in the supernatant after centrifugation to approximately 1:1, it was possible to prepare a capsule content liquid composition containing up to 4.4% dissolved sesamin by weight, even when using MCT, an oil with a low ability to dissolve sesamin.

(Comparative Example 1)
The same treatment as in Example 1 was carried out except that the sesame extract and MCT were mixed in the composition ratio shown in Table 6.
The compositions (theoretical values) of sesamin and sesamolin contained in the blended composition at the time of blending are shown in Table 6, and the concentrations of sesamin and sesamolin contained in the supernatant (measured by LC) are shown in Table 7.

Even when sesame extract products A and B, which have a high sesamin content, were used in the blended composition in the same amount as sesame extract product C (Reference Example 1), which has a higher sesamin content, when the sesamin:sesamolin concentration ratio in the supernatant after centrifugation was less than 1:1, the MCT was unable to dissolve sesamin at a concentration of 2% by mass or more in the capsule content liquid composition.

As described above, when the sesame extract concentration in the blended composition was 30% by mass or less, the solubility of sesamin in the capsule content liquid composition could be increased to 19.0% by mass by using clove oil as a solvent, regardless of the sesamin:sesamolin concentration ratio (Table 3).

Sesamin and sesamolin are considered to mutually improve the solubility. The total of sesamin and sesamolin concentrations is higher when the ratio of sesamin to sesamolin in the centrifugal supernatant is about 1:1 than when the ratio is about 3.5:1. For example, the total concentrations of sesamin and sesamolin in the centrifugal supernatants C-2, C-5, and C-8 are 17.1%, 20.9%, and 24.3%, respectively (Table 3), while the total concentrations of sesamin and sesamolin in the centrifugal supernatants C-3, C-6, and C-9 are 16.4%, 20.6%, and 25.2%, respectively (Table 3). Therefore, when the concentration of sesame extract in the blended composition is increased to more than 30% by mass, the total concentration of sesamin and sesamolin is higher when the ratio of sesamin to sesamolin in the centrifugal supernatant is about 1:1, and as a result, it can be estimated that the concentration of sesamin in the centrifugal supernatant is also higher.
The above inference is also suggested by the results of the MCT solubility tests (Reference Example 1 and Comparative Example 1), and it is believed that when sesame extract is dissolved in clove oil at a higher concentration, the solubility of sesamin increases when the concentration of sesamolin in the capsule content liquid composition is higher.

(Comparative Example 2)
The same treatment as in Example 1 was carried out except that sesame extract C and rice germ oil were mixed in the composition ratio shown in Table 8.
The compositions (theoretical values) of sesamin and sesamolin contained in the blended composition at the time of blending are shown in Table 8, and the concentrations of sesamin and sesamolin contained in the supernatant (measured by LC) are shown in Table 9.

Rice germ oil was unable to dissolve sesamin in the capsule content liquid composition at a concentration of 0.08% by mass or more.

(Comparative Example 3)
The same treatment as in Example 1 was carried out except that sesame extract C and sesame oil were mixed in the composition ratio shown in Table 10.
The compositions (theoretical values) of sesamin and sesamolin contained in the blended composition at the time of blending are shown in Table 10, and the concentrations of sesamin and sesamolin contained in the supernatant (measured by LC) are shown in Table 11.

Sesame oil was unable to dissolve sesamin in the capsule content liquid composition at a concentration of 0.2% by mass or more.

(Example 2)
(Transparency evaluation test)

1. Evaluation method for transparency evaluation test The soft capsule content liquid compositions C-3, C-6, and C-9 prepared in the solubility test were subjected to ultraviolet-visible absorbance measurement using an ultraviolet-visible spectrophotometer (HITACHI U-3900 Spectrophotometer) to quantitatively determine the transparency of the solution and solvent in the vial. The transparency was evaluated using the transmittance at 750 nm.
The transparency of these soft capsule content liquid compositions was evaluated using the following four-level scale (◎, ◯, △, ×).

◎: 750 nm transmittance is 95% or more. ◯: 750 nm transmittance is 80% or more. △: 750 nm transmittance is 60% or more but less than 80%. ×: 750 nm transmittance is less than 60%.

The results of the transparency test for the soft capsule liquid compositions C-3, C-6, and C-9 prepared in the solubility test are shown in Table 12.

Next, as control products, the liquid contents of a soft capsule with sesamin dissolved therein (Suntory Ltd., Sesamin EX) (Control Example 1) and the liquid contents of a soft capsule with sesamin dissolved therein (Kobayashi Pharmaceutical Co., Ltd., Kobayashi Pharmaceutical's Sesamin) (Control Example 2) were used, and the transmittance of these liquid contents to visible light at 750 nm was measured, and a transparency evaluation was performed using the same criteria as above. The results are shown in Table 13.

The capsule content liquid compositions C-3, C-6, and C-9 of the present invention dissolved sesamin in the capsule content liquid at a higher concentration than Suntory's Sesamin EX and Kobayashi Pharmaceutical's Sesamin, and showed the same level of transparency as these control products.

(Comparative Example 4)
The transmittance of visible light at 750 nm through the liquid contained in a soft capsule ("Sesamin Capsule" by Organland Co., Ltd.), in which sesamin is emulsified and suspended in a base oil using an emulsifier and beeswax, was not measurable.

By using this invention, it is possible to increase the amount of ingredients ingested per capsule while maintaining the transparency of the capsule content liquid, thereby reducing the number of capsules to be taken orally. This also reduces the capsule processing costs, thereby reducing production costs.

Claims (6)

Contains lignan compounds including sesamin and sesamolin dissolved in clove oil .
the concentration ratio of sesamin:sesamolin dissolved in the composition is 78:22 to 50:50;
The composition has a total concentration of sesamin and sesamolin dissolved therein of 16.4 to 25.2 mass % . The composition of claim 1, wherein the concentration of sesamin dissolved in the composition is from 8.3 % to 19.0 % by weight. The composition according to claim 1, which has a transmittance of 80% or more at a wavelength of 750 nm in ultraviolet-visible absorbance measurement. A food or drink comprising the composition according to any one of claims 1 to 3 . The food or drink according to claim 4 , which is in the form of a capsule. A method for increasing the solubility of sesamin in clove oil by dissolving sesamin and sesamolin such that the sesamin:sesamolin concentration ratio is 78:22 to 50:50, so that the total concentration of sesamin and sesamolin dissolved in the clove oil is 16.4 to 25.2 mass%.