JP6086264B2 - Method for producing after-scented silk - Google Patents

Description

The present invention relates to a substantivity sills click manufacturing how consisting inclusive and silk incense ingredients, more particularly, by mixing a fragrance component of silk solution prepared from silk protein, obtained by a predetermined process, incense a substantivity silk comprehensive components, various aroma components stably adsorbed relates to the production how the substantivity sill click excellent in persistence of scent.

  Recently, incense ingredients are widely filled around us. In addition to enjoying the aroma, it is believed that the incense ingredients have a mysterious power to heal mental and physical diseases, and incense ingredients have been commercialized. Regarding the effects of the fragrance itself on the mind and body, in general, aromatic oils containing alcohols, ketones, terpenes, and esters as the main components have a sedative effect, and aromatic oils containing aldehydes, phenols, and fatty acid esters are excited. It is said to have a medicinal action.

  In order to utilize these fragrance ingredients economically and efficiently, it is important to effectively volatilize a small amount of fragrance ingredients over a long period of time. ing.

  In the case of insect biopolymer silk, silkworms fed with mulberry leaves as feed are biosynthesized in the silk gland and stored in the body, and then spun into silk thread. It has been suggested that silk thread is a material that can adsorb the scent component, but the method of applying the scent component is difficult and has many problems to be solved.

  The protein fiber produced by spitting a mature moth larva is the cocoon fiber (silk thread). There are two types of silkworms: a rabbit raised by a farmer and a wild-type wild boar. Silk fibroin (also referred to as silk fibroin fiber) is obtained by removing sericin, an adhesive substance covering the surface of the cocoon fiber, by alkali treatment or the like.

  Wild silk thread is a silk thread that is usually spun by Antheraea pernyi, Antheraea yamamai, Antheraea militta, Antheraea assama, Eli and Shinju. Say.

  To prepare silk fibroin aqueous solution from silk protein fiber, raw silk and silk thread derived from rabbit or wild silkworm are used, and when these raw silk or silk thread is boiled with alkaline aqueous solution such as sodium carbonate, it is a gelatinous protein that covers the silk thread surface. Some sericin can be removed and silk fibroin fibers can be prepared. The silk fibroin fiber thus prepared can be heated and dissolved with a concentrated neutral salt aqueous solution, placed in a cellulose dialysis membrane, and replaced with water to prepare a pure silk fibroin aqueous solution.

  When this silk fibroin aqueous solution is dried and solidified on a substrate film such as a polyethylene film, a transparent silk fibroin film (silk film) can be formed. Further, when a silk fibroin aqueous solution is freeze-dried, silk sponge (silk porous material) or silk powder can be produced.

  In order to dissolve silk fibroin fibers derived from rabbits, generally known neutral salts such as calcium chloride, calcium nitrate, and lithium bromide can be used. In order to increase the solubility of the silk thread and produce silk fibroin fibers close to the unmodified state, lithium bromide of lithium ion having high solubility is particularly preferably used.

  In order to prepare silk fibroin fibers derived from wild silkworms, in the case of wild silk threads obtained from silkworms or tengu, for example, these wild silk threads are immersed in an aqueous solution of 0.1% sodium peroxide that is 50 times the weight of the silk thread. However, it is necessary to remove sericin beforehand by treatment at 98 ° C. for 1 hour. A wild silk silk fibroin aqueous solution can be prepared by dissolving wild silk silk fibroin fiber from which sericin has been removed with a neutral salt having high solubility such as lithium thiocyanate, and then dialyzing it with a pure dialysis membrane.

  Rabbit silk and wild silk are the same silk protein derived from silkworms, but the chemical structure is completely different and the physicochemical properties of silk are also greatly different. Wide application development as industrial materials utilizing these characteristics is strongly desired.

  Since silk has a uniform molecular structure and excellent crystallinity, it is difficult to fill the fragrance component into the fiber. Therefore, a method of attaching the scent component directly to the surface of the sample or attaching the scent component as a small-shaped bead containing the scent component may be considered. However, there is a problem with the durability of the scented fiber produced by such a method, and the scented effect is weakened during the storage period or the scent component is volatilized by physical stimulation, and the scented effect is attenuated. The problem arises.

  Hereinafter, the utility of the incense component will be described. As described above, the incense component can be a psychological and mental stimulus for a person. German psychiatrist Terenbach explains that olfaction, like taste, is a “sense of mediating human encounters in the deepest part”. It is known that odor (fragrance) affects not only human physiology and mental activity, but also various functions of the body. Currently, research using electroencephalograms and magnetoencephalograms is progressing in various fields. However, it was around 1980 that the research to scientifically prove the function and effectiveness of odors began.

  Next, the names of the component compounds and their pharmacological effects will be described for the incense components around us.

  Plants that are used as ingredients for incense ingredients often have the scientific name officinalis (medicinal), and the aromatic oils obtained from these plants not only enjoy the scent but also have a mysterious power to heal mental and physical diseases. It has been believed for a long time. Regarding the effects of the scent itself on the mind and body, as described above, in general, aromatic oils containing alcohol, ketone, terpene, and ester as main components have sedative effects, and include aldehydes, phenols, and fatty acid esters. Aromatic oils are said to have stimulant effects. (For example, see http://www.jffma-jp.org/learning/base/bionomy.html)

  Incense ingredients contained in the fragrance are expected to have the following effects. That is, (1) rests feelings and nerves, (2) calms and brightens the mind, (3) warms the mind and body, and stabilizes the emotion.

  Let's look at examples of incense ingredients around us. In the present specification, for example, it is not a description of ingredient flavors of natural fruit juice such as rosemary oil, strawberry (Strawberry), or vanilla oil, but as a main component, cineol (Cineol), furaneol (Furaneol) In some cases, the ingredient name is Vanillin.

(1) Cineol An oily fragrance component of rosemary oil. Also called eucalyptol or 8,1-cineole. It is familiar as a cooking herb. It is said to be effective in eliminating stagnation of moisture and tightening skin, and is sometimes used for beauty treatment. The essential oil of rosemary oil is roughly divided into three chemotypes: cineol, verbenone, and camphora. Increased memory and concentration by stimulating action and brain clearing action, as well as arousal and appetite suppression (for example, “Small Encyclopedia” by Yozo Watanabe, http: // aroma-guide. net / essential / rosemary-cineol.html).

(2) Furaneol Presents in a solid state with an incense component such as strawberry. Furaneol is the name of 2,5-dimethyl-4-hydroxy-3 (2H) -furanone (4-hydroxy-2,5-dimethyl-3 (2H) -furanone). The concentration used in the final product is about 5.0 to 10.0 ppm (synthetic fragrance chemistry and product knowledge, Motokazu Into, Chemical Industry Daily, 2005). Strawberry is a plant of the Rosaceae family, and the scent of roses generally has a pharmacological effect that brightens the mood and has a sedative effect (for example, http://www.google.com/patents/ WO2011055789A1? Cl = en; see http://www.paradise-g.jp/ichigo.php).

(3) Vanillin An oily form with a scent component of vanilla oil. It is said to have a relaxing effect, relieve anger, frustration and tension, make you happy, and be effective for insomnia (eg http://www.timeless-edition.com/archives/ 2959; see http://www.deodor.co.jp/kaori/effect.html).

(4) Menthol
It is an incense component contained in mint and is said to have the effect of awakening sleepiness, clarifying consciousness, and calming the heart (for example, see http://matome.naver.jp/odai/2133777182 036211101) ).

(5) Limonene
It is an incense component contained in lemons and is contained in citrus fruits in addition to lemons. It is said to refresh the mood, clarify consciousness, and improve understanding and concentration (see, for example, http://www.aromatherapy-style.net/Citrus-limon.html).

(6) Linalool
It is an incense ingredient contained in lavender and is effective for relaxation, restful sleep, relaxation of tension and stress (for example, see http://www.gosetsumei.com/lavender.html).

(7) Cinnam aldehyde
Incense component of cinnamon. It is said that it is uplifting, uplifting, stimulating, and energizing a fatigued / debilitated heart (eg http://www.aromatherapy-style.net/cinnamomum-zeylanicum.html) ).

  Conventionally, the following techniques are known for the scented fiber. A technique is known in which a fiber is scented by bringing a fragrance into contact with a fragrant fiber using a fragrance imparting agent containing a predetermined compound (see, for example, Patent Document 1). Specifically, Patent Document 1 discloses a durable fragrance treatment method in which a treatment liquid containing an inorganic porous powder adsorbing a fragrance and a silicone compound is applied to a fiber structure, followed by drying and heat treatment. Has been. However, the method of Patent Document 1 has problems that should be improved in the sustained release property of the fragrance component from the fiber containing the fragrance, the stable durability, the washing durability, and the like.

  In addition, as a technique for adsorbing perfume to the fiber, in order to provide a fiber with excellent scent release properties, stable sustainability and durability for washing, etc. A fiber and a perfume method for the fiber have been proposed (see, for example, Patent Document 2). That is, an invention relating to a residual fragrance imparting agent for a fiber containing an organosiliconate and water, a residual fragrant fiber containing a cross-linked polyorganosiloxane containing a silanol group, a method for producing the same, and a method for flavoring the fiber are disclosed. However, chemical skills and knowledge have not been proposed for the method of flavoring residual fiber.

  Furthermore, regarding the residual fragrant fiber material, can the fragrance be stably supported, the fragrance component does not decay over time, or the fragrance may change in quality or the fragrance may vaporize to cause structural destruction. No prior literature is found regarding technology that satisfies the above.

JP-A-1-246478 JP 2008-223156 A

  As mentioned above, there are various scents around us, and sometimes there are scents that give people psychological comfort. In the food field, foods and seasonings that contain a fragrance are commercially available. Since fragrance ingredients and herbal ingredients derived from plants generally have a high vapor pressure, there is a problem that if the fragrance ingredients volatilize during storage when stored in a standard state for a long time, the residual fragrance effect is attenuated.

  Since many scent components are volatile, storing the scent component for a long period of time reduces the scent component and reduces the amount of volatilization, thereby reducing the intensity of the scent. Need to be developed. For this purpose, it is inconvenient for a material that adsorbs only the surface of the material when the scent component is adsorbed to the material. Usually, a material in which an incense component is adsorbed only on the surface often decreases the volatilization amount of the incense component for a long time or causes an adverse effect of the incense component. That is, if a scent component is adsorbed on ordinary organic fibers, the fiber may be discolored due to the scent component or the physical properties of the fiber may be deteriorated. It was. In addition, when organic fibers containing an incense component are washed, the incense component is detached from the fiber, which is not suitable as an inclusive material for the incense component.

  In addition, when trying to produce an ideal residual scented material, there is a problem in that the manufacturing cost becomes high. In general, no matter what material the fragrance component is adsorbed, the fragrance component volatilizes over time and the intensity of the fragrance decreases. Therefore, when a scent component is required, the appearance of a wrapping material having a function of enhancing the strength of the scent component once included in the material, or a function that increases the volatilization amount of the scent component at a required time. The appearance of the remaining scented material is strongly desired. However, there was a problem that an effective aftercense material could not be found.

  In the present invention, lyophilization is required to remove water from the silk fibroin aqueous solution to which the fragrance component has been added. In this case, two steps of preliminary freezing and lyophilization are necessary. However, an economical and efficient temperature and pre-freezing time for pre-freezing a predetermined concentration of silk fibroin aqueous solution added with a fragrance component has not been clarified in the prior art. Further, even in the freeze-drying process following the pre-freezing, an economical and efficient freeze-drying temperature and freeze-drying time have not been clarified in the prior art. It is necessary to clarify such optimum conditions, and there is still no known simple method for detecting useful solvents and novel fragrance components.

  A gas chromatograph (GC) or a gas chromatograph-mass spectrometer (GC-MS) is convenient for investigating whether a scent component is volatilized or gradually released from a material containing a scent component easily vaporized. . In other words, if a substance in a material is separated by GC, and it is guided directly to the MS from the outlet of GC and a plurality of separated components are measured, a small amount of components can be analyzed conveniently and highly sensitively without waste. it can. If a detector with high measurement sensitivity is used, even commercially available products can be detected to the order of several tens of fg / s (femtogram per second), and are widely used as trace analysis techniques in various scientific fields.

  As in the present invention, in order to measure the fragrance component vaporized from the sample containing the fragrance component, GC, a container for collecting the fragrance component to be vaporized prior to GC-MS measurement, and a device for taking out the fragrance component to be vaporized , And whether a column tube filled with a filler for adsorbing the fragrance component is necessary, and what solvent is used to remove the gas once adsorbed to the column filler, ie, vaporize from the sample It is necessary to search for the optimum conditions for measuring the scent component by GC-MS. However, the technology regarding such conditions is not known.

  Since the above-described measurement for quantifying the fragrance component that is easily vaporized requires skillful chemical knowledge, it is desired that there is a simple method that complements the GC-MS measurement. Sensory evaluation that evaluates incense components based on human sense of smell is another option. In the present invention, the application of such a method has been studied with a view point in mind, but there is a problem that there are many unclear parts.

  An object of the present invention is to solve the above-mentioned problems of the prior art, and it is possible to adsorb a fragrance component to silk fibroin, and furthermore, a scented silk excellent in sustained release and sustainability of the adsorbed fragrance component and An object of the present invention is to provide a silk composite, and a method for perfusing silk and silk composite.

  The problem of the present invention is also to solve the above-mentioned problems of the prior art, and to evaluate a method for enhancing a scent component that evaporates from a solid silk containing a scent component or a scent component that evaporates from a sample after a certain period of time. The object is to provide a scientific method, a method for detecting a scent component, and a technique for producing a scented silk.

  The inventors of the present invention have been developing a new technology for utilizing a silk fibroin aqueous solution in which rabbit or wild silk yarn is dissolved. Development of materials that economically and efficiently produce residual scented silk by pre-frozen silk fibroin aqueous solution containing scented ingredients, followed by freeze-drying, and repeated research to evaluate the amount of scented ingredients that evaporate over time We have been conducting research on the evaluation of incense ingredients using a wide range of technologies.

  The present inventors also describe how to mix an incense component into an aqueous silk fibroin solution, how to dry the silk fibroin aqueous solution, how to evaluate the incense component economically and effectively, etc. As a result, the evaluation method of a fragrance | flavor component and the reinforcement | strengthening method of the fragrance | flavor component volatilized from solid dry silk were established.

  The present inventors have further noted that natural proteins effectively adsorb gas having a small molecular weight such as a fragrance component, and silk powder or silk sponge (silk porous body) obtained from silk fibroin fiber is used. It has been found that it is effective to contain a fragrance component, and that the amount of vaporization of the fragrance component once adsorbed is increased by adding water.

  As the silk fibroin aqueous solution, a silk aqueous solution derived from rabbits and wild silkworms can be used.

According to the scented silk obtained by the method for producing a scented silk of the present invention, a scent component is added to a silk fibroin aqueous solution, pre-frozen, and then freeze-dried, whereby the molecular structure of the silk having a random coil shape As a result, it is possible to prevent the incense component from volatilizing even after a long period of time. The strength can be prevented from decreasing. Moreover, since silk powder, silk sponge, etc. are couple | bonded through a strong interaction with a fragrance | flavor component, washing durability is also excellent.

According to the scented silk obtained by the method for producing a scented silk of the present invention, a scent component is added to a silk fibroin aqueous solution of silk protein, this aqueous solution is pre-frozen, and then freeze-dried, in a silk fibroin aqueous solution. By removing the moisture, the incense component remains stably in the silk fibroin for a long period of time, and as a result, the incense component can be stably stored for a long period of time.

  Silk proteins that can be used include silk fibroin fibers derived from rabbits or wild silkworms. The function of the residual scented silk is enhanced by using rabbit silk and wild silk, especially wild silk silk as the material of the residual scented silk.

Incense ingredients that can be used in the present invention is not particularly limited, among the incense components described above, for example, rosemary oil, rose oil, scan Toro berry flavor can be preferably used.

  The present inventors have found that silk fibroin in the form of powder or sponge of the fiber is superior to the silk protein fiber (silk fibroin fiber) itself as an adsorption form. Silk fibroin, which is an adsorbed form such as a powder form or a sponge state rather than a fiber form, can be produced by adding a perfume component to silk fibroin in an aqueous solution, pre-freezing, and then drying and freezing. In addition, when silk fibroin aqueous solution is freeze-dried, silk sponge or silk powder can be produced.

In view of the above, the detection method of the incense ingredient substantivity silk obtained substantivity silk and the manufacturing method sequence obtained by the present invention is as follows.

The scented silk obtained by the present invention is a pre-frozen and subsequently freeze-dried incense component, silk fibroin derived from rabbit, silk fibroin derived from silkworm, silk fibroin derived from rabbit and silk fibroin derived from silkworm ing from solid dry silk and a silk fibroin selected from the complex.

The scented silk obtained by the present invention is also pre-frozen and subsequently freeze-dried, and the scent component, silk fibroin derived from rabbit, silk fibroin derived from silkworm, silk fibroin derived from rabbit and silk fibroin derived from silkworm of the substantivity silk consisting of solid dry silk and a silk fibroin selected from the complex, and the aqueous, ing and preferably added prior to use.

In substantivity silk obtained by the present invention, the mixture of the aroma component and silk fibroin formed by solidified frozen by preliminary freezing, ing lyophilized as moisture contained therein is sublimated.

Method for producing a substantivity silk of the present invention, rosemary oil, rose oil, and the fragrance component selected from the strawberry flavor, at a temperature of the silk solution silk Fiburoi emissions from silkworm -10 ℃ ~-80 ℃ , Pre-freeze for 9 to 60 minutes , and then solidify completely, and then the moisture contained in the fragrance component and the silk fibroin derived from the rabbit are completely frozen by this preliminary freezing. lyophilized at a -50 ° C. so as to sublime, can be prepared a substantivity silk consisting of solid dry silk and a silk fibroin derived from the aroma component and pedigree silkworms, and substantivity silk produced is It is characterized by being added with water at the time of use .
The dried silk derived from rabbit has a random coil-like molecular structure.

Method for producing a substantivity silk of the present invention also includes rosemary oil, rose oil, and the fragrance component selected from the strawberry flavor, wild silkworm derived silk fibroin, and silk fibroin and silk fibroin derived from the wild silkworm derived silkworm The silk fibroin silk aqueous solution selected from the above composites was pre-frozen at a temperature of −10 ° C. to −80 ° C. for 9 minutes to 60 minutes , and then frozen and solidified by this preliminary freezing. freeze-dried at -50 ° C. so as to completely sublime the water contained in the complex of silk fibroin from silk fibroin and pedigree silkworm derived silk fibroin or wild silkworm-derived components and wild silkworms, the incense ingredient including solid and the silk fibroin from wild silkworm or the complex of the flavoring component and pedigree silkworm derived silk fibroin and wild silkworm derived silk fibroin of To produce the substantivity silk consisting of dry silk from Jo of wild silkworm or to produce a substantivity silk consisting of a complex between the solid dried silk and solid dry silk from silkworm-derived wild silkworms, The manufactured scented silk is characterized by being added with water at the time of use .

In the method for producing a scented silk of the present invention, the dried silk derived from rabbit has a random coil-like molecular structure, and the dried silk derived from wild silk has a molecular structure of a random coil and an α helix. characterized in that it have a.
In the method for producing after-scented silk of the present invention, the freeze-drying is carried out at −50 ° C. for 3 hours to 15 hours.

In the method for manufacturing the substantivity silk, wherein the concentration of the silk solution of silk fibroin is 0.5 to 1.5 wt%.

In the method for manufacturing the substantivity silk, solid dry silk obtained after freeze drying, wherein the drying silk der Rukoto powdered dry silk or porous body.

Oite the manufacture how the substantivity silk described above, for example, performed over 10 minutes at -50 ° C. in pre-freezing, preferably carried out over 10 hours at -50 ° C. in freeze-dried, and the incense ingredient , rosemary oil, rose oil, it is preferred that you and strawberry flavor. If the preliminary freezing temperature and time and the lyophilization temperature and time are used, the intended purpose can be achieved.

  According to the present invention, silk fibroin derived from rabbit or wild silk, or composite silk fibroin composed of rabbit silk fibroin and wild silk fibroin, incense is added to solid dry silk made of silk powder or silk sponge (porous body). Since the ingredients are supported, the incense ingredients are stably and strongly bound to the silk fibroin solid dry silk, so the incense ingredients will not deteriorate or vaporize even if stored for a long period of time. Moreover, it is excellent in washing durability, and the scent component is stably contained in silk fibroin solid dry silk. That is, the fragrance component does not change even during the period of storing the silk fibroin solid dried silk containing the fragrance component. By adding a solvent such as water to a solid dry silk of silk fibroin containing an incense component, the volatilization amount of the incense component from the silk fibroin containing the incense component can be enhanced. Further, according to the present invention, the silk film containing the fragrance component also has a residual fragrance, and the evaluation value of the fragrance increases even when water is added compared to before adding water.

The graph which shows the relationship between preliminary freezing temperature and preliminary freezing time. The gas chromatogram of rosemary oil used by this invention. The gas chromatogram of furaneol used by this invention. The gas chromatogram of vanilla oil used by this invention. The side view of the vaporization apparatus used by this invention. The graph which shows the result of the sustained release property (comparison by peak area) of cineole from the rosemary oil containing dry silk in Table 9.

  According to the present invention, the solid dry silk that can be used to include the incense component consists of silk fibroin derived from rabbit or wild silkworm. As a raw material for producing silk fibroin, silk thread derived from rabbit or wild silk can be used conveniently. As silk fibroin for silk thread, silk protein derived from Bombyx mori larvae bred by farmers and close relatives of silkworm larvae. Alternatively, wild silk fibroin, as described above, for example, from larvae such as Antheraea pernyi, Antheraea yamamai, Tather moth (Antheraea militta), Muga moth (Antheraea assama), Eli moth, Shinju moth, etc. The resulting silk fibroin can be used. As a solid dry silk, a silk-like silk fibroin derived from a rabbit and a silk fibroin derived from a silkworm are pre-frozen and then freeze-dried to obtain a powdered silk derived from a rabbit. It becomes a residual fragrance material superior to the residual fragrance property.

  A method for producing a silk fibroin aqueous solution, powdered silk fibroin (also referred to as silk powder), or silk fibroin film from rabbit silk yarn or wild silk yarn is as follows. In addition, it can form in a powder form, a film | membrane form, and a porous body by changing the evaporation rate and preparation conditions of silk fibroin aqueous solution according to a known method.

Preparation of silk fibroin aqueous solution, silk powder and silk membrane derived from rabbits:
The silkworm silk spun from the silkworm is boiled in an alkaline aqueous solution of neutral salt such as sodium carbonate, and sericin is removed to produce silk fibroin fiber which is the main body of silkworm silk.

  Next, this silk fibroin fiber is dissolved in a concentrated neutral salt aqueous solution and heated to obtain a silk fibroin aqueous solution. Since this silk fibroin aqueous solution contains many ions based on neutral salts in addition to silk fibroin, put it in a cellulose dialysis membrane, tie both ends with sewing thread, and add 2 to room temperature tap water or pure water. A pure rabbit silk fibroin aqueous solution can be produced by performing dialysis treatment for a predetermined time after 5 days. By evaporating a part of water from this silk fibroin aqueous solution or adding water, silk fibroin aqueous solutions having different concentrations can be prepared. This preparation method will be described in more detail below.

  Silk powder can be manufactured by freeze-drying the silkworm silk fibroin aqueous solution thus prepared. Alternatively, a silk membrane can be prepared by spreading a rabbit silk fibroin aqueous solution on a substrate such as a polyethylene membrane and evaporating to dryness at room temperature.

  In addition, when preparing a silkworm silk fibroin aqueous solution by dissolving silkworm silk protein fiber (silk thread), the silk thread is usually added to a concentrated neutral salt aqueous solution such as calcium chloride, calcium nitrate, lithium bromide or lithium thiocyanate. It is done by putting and heating. The neutral salt concentration is about 5 to 9M, and the dissolution temperature is about 25 to 70 ° C, preferably about 25 to 60 ° C. When the melting temperature is higher than 70 ° C., the molecular weight of the silk protein is lowered, and the high molecularity of the material is lost. As a result, there is a risk that the moldability is deteriorated. The dissolution time is preferably set to about 1 to 20 minutes. What dissolves silkworm silk protein fibers satisfactorily is a lithium salt excellent in dissolving power of silkworm fibroin fibers among neutral salts, and lithium bromide is usually preferred. For example, in the case of a lithium bromide aqueous solution of 8M or more, preferably 8.5M or more, the silkworm silk protein fiber is dissolved by the treatment at 55 ° C. or more for 15 minutes or more.

Preparation of silk fibroin aqueous solution derived from wild silkworm:
In order to prepare a field silk silk fibroin aqueous solution from field silk such as silkworm silk or tengu silk thread, first, the field silk thread is immersed in a predetermined amount of sodium peroxide aqueous solution with respect to the weight of the silk thread and boiled for a predetermined time, The prepared wild silk fibroin fiber is dissolved in a highly soluble neutral salt solution. The resulting aqueous solution is then dialyzed in the same manner as in the case of silkworm silk to prepare a pure wild silk fibroin aqueous solution. This preparation method will be described in more detail below.

  In order to dissolve the wild silk yarn and prepare the wild silk silk fibroin aqueous solution, it is necessary to remove the silk sericin covering the surface of the wild silk yarn by a method different from the scouring of the silkworm silk sericin. This is because in addition to sericin, tannin also adheres to the surface of wild silk thread, and sericin is insolubilized by the crosslinking action of tannin. In order to remove these sericin and tannin, for example, it is necessary to immerse a wild silk thread in a 0.1% sodium peroxide aqueous solution of about 50 times the weight of the silk thread weight and boil at 98 ° C., for example, for 1 hour. is there. Wild silk silk fibroin fiber from which sericin and tannin have been removed in advance is dissolved in an aqueous neutral salt solution having high solubility such as lithium thiocyanate. This neutral silk fibroin fiber neutral salt aqueous solution is put in a cellulose dialysis membrane, both ends are sewn with sewing thread, put in room temperature tap water or pure water for 2 to 5 days, and pure ions are removed by completely removing lithium ions. Wild silk fibroin aqueous solution can be prepared.

  By treating the aqueous silkworm fibroin solution thus prepared in the same manner as the silkworm silk fibroin aqueous solution, silkworm-derived silk powder, silk membrane, and the like can be prepared.

Preparation of mixed silk fibroin aqueous solution of rabbit silk fibroin and wild silk fibroin:
A mixed silk fibroin aqueous solution of rabbit silk fibroin and wild silk fibroin prepared by combining the rabbit silk fibroin aqueous solution and wild silk fibroin aqueous solution prepared as described above in an arbitrary volume (for example, 50:50) is provided. A mixed silk fibroin aqueous solution containing both can be prepared by placing in a fixed beaker and carefully stirring and mixing with a glass rod so that the aqueous solution does not gel.

  By treating the mixed silk fibroin aqueous solution thus prepared in the same manner as the case of the silkworm silk fibroin aqueous solution, it is possible to prepare a silk powder, a silk membrane, and the like composed of a composite of rabbit silk fibroin and wild silk fibroin.

  The concentration of the above-described aqueous silkworm fibroin solution and wild silk fibroin aqueous solution is about 0.1 to 6% by weight in order to produce powdery or porous dry silk by preliminary freezing and freeze-drying, respectively. 0.4 to 4% by weight is preferable. In the case of a mixed silk fibroin aqueous solution, of course, aqueous solutions having different concentrations may be used as appropriate. When the concentration of the silk fibroin aqueous solution is less than 0.4% by weight, the amount of silk fibroin contained in the aqueous solution is small, and there is a large amount of the aqueous solution to be evaporated. When the amount is less than 1% by weight, the tendency is further increased. Therefore, it is not efficient and economical in producing a sample. When the concentration exceeds 4% by weight, when handling silk fibroin aqueous solution, if fibroin is mechanically stimulated, the risk of solidification increases, making it difficult to produce finely powdered silk or porous silk. There is a tendency, and when it exceeds 6% by weight, the tendency further increases. Therefore, 0.4 to 4% by weight is particularly preferable. When producing a complex of rabbit silk fibroin and wild silk fibroin, the respective concentrations may be as described above. When the silk fibroin concentration exceeds 6% by weight and becomes 7% by weight, even if the silk fibroin aqueous solution is pre-frozen and freeze-dried, it does not become a powder state with a large surface area, and the solid surface (blocked) or block shape with a small surface area. Fibroin (see Comparative Example 2). As described above, solid powder silk can be formed at 4% by weight or less, and, for example, porous silk can be formed at about 5 to 6% by weight, and bulk silk can be formed at 7% by weight or more.

  From the above-mentioned rabbits, wild silkworms, and mixed silkworms and silkworms derived from silkworms, added a fragrance or a solvent containing a fragrance as a fragrance ingredient, for example, after pre-freezing at −50 ° C. for a predetermined time, When dried for a predetermined time with a freeze dryer set at −50 ° C., dried silk that is a silk fibroin in a solid state containing a fragrance can be produced.

[Aroma evaluation]
After preliminary freezing and freeze-drying, a silk fibroin sample containing a fragrance as a scent component is allowed to stand in the atmosphere for a certain period of time, and the sample is subjected to scent evaluation immediately before the evaluation by the following method. , And gently agitate, let stand for a certain period of time, or after completely dissolving the solid silk fibroin over time, evaluate the fragrance of the fragrance again.

  The fragrance ingredient that can be used in the present invention can be any known fragrance, but it is familiar to cooking around us, such as rosemary oil, rose oil, vanilla oil, strawberry flavor. Can be preferably used.

  A solvent having excellent solubility can be used together with the fragrance component. For example, in the case of rosemary oil, rose oil and vanilla oil, ethanol can be preferably used, and in the case of strawberry flavor, water other than ethanol can be preferably used.

In this invention, sensory evaluation was performed as follows about fragrance.
With respect to the solid aftertaste silk of the present invention, when a predetermined time elapses after the production, four evaluators determine the scent in the vial containing the aftertaste silk as the four-step functional aroma described below. Evaluation was performed according to the evaluation criteria. Furthermore, after evaluating the fragrance, a predetermined amount (for example, 10 mL) of distilled water was added to the vial, and after stirring well, the fragrance was evaluated again in accordance with the functional odor evaluation criteria.

  In addition, the evaluation of aroma was performed by four people in four stages, and a test of significance (one-factor analysis of variance (in-subject plan)) was performed by quantifying the evaluation. However, when rational numbers could not be determined, it was decided to add or reduce 0.5. That is, for example, when neither “1” nor “2” can be determined, “1.5” is set.

  In the present invention, as described above, any fibrous silk protein derived from silkworms such as rabbits and wild silkworms from which sericin is removed in the refining process of the silk thread can be used regardless of the type.

  Next, the present invention will be described in more detail with reference to examples. The present invention is not limited by these examples. In the examples, “%” is “wt%”. Before describing specific examples, first, reagents and measurement conditions used in the following examples will be described.

[Production of silk fibroin fiber derived from rabbits]
Rabbit silk thread was placed in a 4% strength aqueous sodium hydrogen carbonate solution and boiled for 1 hour for refining, and silk sericin was removed to produce silk fibroin fiber.

[Preparation of silk fibroin aqueous solution derived from rabbit]
Silk fibroin fiber by adding 2.5 g of scoured silk fibroin fiber to a beaker containing 65 g of calcium chloride and 60 mL of ion-exchanged water, and heat-treating for 5 minutes with a gas burner using a wire mesh with glass wool. Was completely dissolved. This was placed in a dialysis membrane made of cellulose (imported and sold by Aedia Co., Ltd., product number UC36-32-100, fractional molecular weight MWCO is 14,000) and replaced with tap water for 2 days, and then with ion-exchanged water for 1 day. Replaced. By adding ion exchange water to the silk fibroin aqueous solution remaining in the dialysis membrane, a silk fibroin aqueous solution adjusted to a desired concentration could be obtained.

[Incense ingredient]
Rosemary oil: Trade name: Rosemary essential oil; Living Wooden Rosemary Cineol Co., Ltd .; Lot # 522 (hereinafter abbreviated as rosemary oil) was used. The main ingredient is cineol.

  Strawberry flavor: In Example 3, a filtrate obtained by filtering cocoons well ground in a mortar through a filter and removing impurities was used. This major component is furaneol. In and after Example 5, furaneol (manufactured by Sigma Aldrich Japan, 2.5-dimethyl-4-hydroxy-3 (2H) -furanone, Lot # STBC7913V) was used.

  Vanilla oil: Vanilla essence (trade name: Essence vanilla; manufactured by Meidi-ya; Lot # 2015.522) was used. This major component is vanillin.

  Rose oil: Rose essential oil (trade name: Maggie Tisserand, Essential Oil for Aromatherapy; hereinafter abbreviated as rose oil) was used. The main component is phenethyl alcohol. This essential oil is a trade name and does not mean the shape of the incense component.

[Used silk fibroin aqueous solution]
In the examples, the silk fibroin aqueous solution derived from the above-mentioned rabbit (hereinafter referred to as “rabbit silk-derived silk aqueous solution”), the silk fibroin aqueous solution derived from the wild silkworm prepared according to the following examples, as the silk fibroin aqueous solution. A mixed silk fibroin aqueous solution derived from rabbit and wild silkworm and a commercially available aqueous solution of powdered silk (hydrolyzed silk; manufactured by Ichimaru Falcos; product name: Edible Silk, Lot # 132851) were used.

[freeze drying]
A scent component solution having a predetermined concentration was added to a silk fibroin aqueous solution having a predetermined concentration. In the case of strawberry flavor, this solution was diluted with water, and in the case of other flavorings and oils, 1 drop (about 0.05 g) of the stock solution was prepared to 20-50% with distilled water. It is appropriately diluted with an ethanol aqueous solution. Place it in a 20 mL vial with a rubber stopper (manufactured by Marum Vacuum Vial, No. 5, 84-1354), and place this vial in a pre-freezer (EYELA, PFR-1000) at -50 ° C for a specified time. (For example, for 20 minutes) The silk fibroin aqueous solution was frozen (pre-freezing is a treatment for completely solidifying the silk in the aqueous solution state). Subsequently, the silk fibroin aqueous solution was freeze-dried by placing it in a freeze-dryer (EYELA, FDU-1200) set at −50 ° C. and allowing to stand for a predetermined time (for example, 15 hours). This is a process of completely sublimating and removing water in the silk solids once solidified). In this case, after completely freezing by preliminary freezing, lyophilization is carried out at a normal temperature and time for removing water in the sample. In other words, the fragrance component and silk fibroin, which are completely frozen and solidified by preliminary freezing, are freeze-dried so that the water contained therein is sublimated. The desired solid dried silk can be produced through lyophilization at the normal pressure reduction described below.

The used pre-freezing tank and freeze-dryer (manufacturer name, standard, etc.), their use time, pre-freezing temperature and freeze-drying temperature, used glass container, etc. are as follows.
Pre-freezing device (tank): EYELA, PFR-1000, operating temperature is -50 ° C, used for a predetermined time just before setting in freeze dryer.
Freeze dryer: EYELA, FDU-1200, operating temperature is -50 ° C, used for a predetermined time.
20 mL capacity glass container and its lid (rubber stopper): No.5, 84-1354, manufactured by Marum Vacuum Vial
Silica gel of desiccant: Sansho (trade name: CD: 59-1266)
Dialysis membrane: Import sales agency: Adia Co., Ltd., product number UC36-32-100, permeable membrane weight (MWCO) of dialysis membrane is 14,000.
The degree of vacuum of the freeze dryer used in the examples was 30 Pa or less. If the degree of vacuum of the specifications of conventionally known freeze dryers, there is no problem in producing silk powder. Satisfactory results can be obtained with general freeze dryer specifications (0.1 Pa-110 kPa). For example, the degree of reduced pressure in freeze-drying is usually 110 kPa or less, desirably 30 Pa or less.

[Aroma sensory evaluation method]
As described above, sensory evaluation of fragrance was performed by four people, and the fragrance was confirmed by bringing the glass container (vial bottle) containing the sample close to the nose with the lid removed. The strength was quantified according to the evaluation criteria of 0.5 increments according to the following criteria.

Sensory evaluation criteria:
Strong aroma: 3
Odor is normal: 2
Fragrance is weak: 1
I do not feel fragrance: 0

  Also, in order to prevent loss of arbitraryness in the evaluation value depending on the order of the samples to be subjected to sensory evaluation, even if the sample has been confirmed fragrance once, the order was changed, the fragrance was confirmed multiple times, and reproducible results It was made to become.

In this example, the optimum conditions for the pre-freezing time and the freeze-drying time were examined.
The silk fibroin aqueous solution used in this example is a silkworm-derived silk aqueous solution having a concentration of 0.5% and a hydrolyzed silk (made by Ichimaru Falcos) aqueous solution. 5 mL of each of these aqueous solutions is put into a vial, and the bottle is put into a pre-freezing apparatus set at −50 ° C., and immediately after pre-freezing at different times (3, 8, 10, 12 minutes), a sample is put. The vial was placed in a freeze dryer set at −50 ° C. and dried under reduced pressure for 3, 8, 10, 15 hours. The dried state of the sample was visually observed.

[Sample condition by preliminary freezing]
When pre-freezing treatment was carried out at −50 ° C. for 3 minutes, the inside of the vial was observed, and in the case of both silkworm-derived silk aqueous solution and hydrolyzed silk aqueous solution, only a part of the sample was frozen. It was in an aqueous solution state. When a sample partially frozen was attached to a freeze dryer at −50 ° C. and a degree of vacuum of 30 Pa, the sample was blown out of the vial, resulting in a foamy sample with a dry state that was easy to handle. Silk powder could not be produced.

  When the preliminary freezing treatment was performed at -50 ° C for 8 minutes, the inside of the vial seemed to be frozen in both cases of the silkworm-derived silk aqueous solution and the hydrolyzed silk aqueous solution, but in the case of 3 minutes When the sample was placed in a freeze dryer under the same conditions as the sample and dried, the sample did not blow out from the vial, but the inner wall of the bottle was slightly wetted to completely remove the moisture in the sample. I understand that I couldn't.

  The inside of the vial when the preliminary freezing treatment is carried out at −50 ° C. for 10 minutes is completely and surely frozen in both cases of the silkworm-derived silk aqueous solution and the hydrolyzed silk aqueous solution. When put in a freeze dryer under the same conditions as the case and dried, all the silks became dry.

  The inside of the vial when the preliminary freezing treatment is performed at −50 ° C. for 20 minutes is completely and surely frozen in both cases of the silkworm-derived silk aqueous solution and the hydrolyzed silk aqueous solution. When placed in a freeze dryer under the same conditions as the case and dried, all the silks became dry. There was no difference from the pre-freezing treatment for 10 minutes.

  From the above experiment, the freezing time when 5 mL of 0.5% silk fibroin aqueous solution is pre-frozen at −50 ° C. is generally 9 minutes or longer, optimally 10 minutes or longer, and then at −50 ° C. for 10 hours. In the case of freeze-drying during this period, it was possible to obtain a silk solid material having a uniform shape and easy to handle. If the preliminary freezing was insufficient, the sample water could not be completely removed when freeze-dried as described above. However, if preliminary freezing was sufficient, the sample moisture could be completely removed when lyophilized. Although there is no restriction | limiting in particular regarding an upper limit time, What is necessary is just to implement about 9-10 minutes or more from a suitable upper limit from the economical viewpoint, for example, when a preliminary freezing temperature is -50 degreeC. That is, it may be carried out for at least the pre-freezing completion time at each pre-freezing temperature (the time required for the pre-freezing to completely freeze and solidify the sample aqueous solution). Next, before finding the optimum conditions for freeze-drying time when freeze-dried at −50 ° C., the relationship between various pre-freezing temperatures and pre-freezing time was examined.

  Before examining the optimum conditions for freeze-drying time, the relationship between the pre-freezing temperature and the pre-freezing time for freezing the sample silk aqueous solution completely and reliably, that is, freezing, was examined. The pre-freezing conditions (temperature and time) vary depending on the amount of the sample silk aqueous solution, but may be any temperature and time at which the sample silk aqueous solution is completely and reliably frozen. For example, the amount of the solution is 2 to 10 mL. In this case, the preliminary freezing temperature may be −10 ° C. to −80 ° C. and the preliminary freezing time may be 60 minutes to 2 minutes. FIG. 1 shows the relationship between the preliminary freezing temperature and the preliminary freezing time. As is clear from FIG. 1, the preliminary freezing time is shorter as the preliminary freezing temperature is lower, and the preliminary freezing time is longer as the preliminary freezing temperature is higher. For example, if the sample silk solution is frozen at −80 ° C. for 2 minutes or more, −50 ° C. for 9 minutes or more, −20 ° C. for 30 minutes or more, and −10 ° C. for 60 minutes or more, the sample silk aqueous solution is completely frozen. Once the sample is pre-frozen and solidified, it is exposed to normal pressure (eg, 0.1 Pa-110 kPa), normal temperature (eg, -50 ° C.), and normal time (eg, 9 hours or more). By freeze-drying, moisture sublimates while solidifying. Freeze-drying may be performed under conditions of temperature and time at which the moisture of the solid sample that has been completely frozen in the preliminary freezing process is sublimated. For example, it is completely frozen and solidified by preliminary freezing. What is necessary is just to implement the silk fibroin on normal conditions so that the water | moisture content contained in this may be sublimated. For example, if lyophilization treatment is performed at −50 ° C. for 10 hours or longer, the sample moisture completely evaporates from the solidified sample in the frozen state. As a result, a desired powdery silk can be produced.

  As described above, the pre-freezing condition is more important than the freeze-drying condition, and it is an indispensable condition that the sample aqueous solution is surely frozen, that is, reliably frozen. For example, it is pre-frozen under the pre-freezing completion time at a predetermined pre-freezing temperature to ensure that the aqueous sample solution is frozen, and then included in the sample by freeze-drying by exposing the solidified sample in a frozen state under a predetermined reduced pressure. The desired moisture can be obtained by sublimating the water content.

  Next, the lyophilization time will be further examined. Pre-freezing was performed on the silk aqueous solution at −50 ° C. for 10 minutes, and freeze-drying was performed after pre-freezing under conditions equal to or longer than the pre-freezing completion time.

  After pre-freezing at -50 ° C. for 10 minutes or more under the condition of the pre-freezing completion time or more, as described below, the moisture contained in the perfume component and silk fibroin that is surely frozen and solidified Was lyophilized to fully sublime.

  Pre-freezing is performed at −50 ° C. for 10 minutes, and the fragrance component and silk fibroin that have been completely frozen and solidified by pre-freezing are freeze-dried (for example, so that the moisture contained therein is sublimated) And freeze-dried at −50 ° C. for 10 hours). For both silkworm-derived silk aqueous solution and hydrolyzed silk aqueous solution, when the vial is taken out from the freeze dryer and allowed to stand, the solid state is maintained, the desired form is prepared, and a silk solid material that is easy to handle can be obtained. did it. If the freezing time is short, even if it is pre-frozen and frozen, the sample will tend to dissolve if left standing, and the intended form is well prepared and a silk solid material that is easy to handle cannot be obtained. there is a possibility. If it was a predetermined time (for example, 9 hours or more), even if it left still, the target form was prepared and the silk solid substance easy to handle could be obtained. The upper limit time is not particularly limited, but is generally 9 hours or longer, and optimally 10 hours or longer from an economical viewpoint.

  From the above experiment, preliminary freezing is performed under the removal condition of the preliminary freezing completion time of, for example, -50 ° C. for 10 minutes or longer, and lyophilization is performed for a predetermined time and temperature (for example, -50 ° C. for 9 to 10 hours or longer). ) In the case of the silkworm-derived silk aqueous solution and the hydrolyzed silk aqueous solution, the silk solid material that does not dissolve even when taken out from the freeze dryer, maintains a solid state, has a desired shape, and is easy to handle. Could get.

  The above experiment is an experiment for clarifying the optimum conditions for the pre-freezing time and the freeze-drying time, and was performed without adding a fragrance component to the sample. Even when the silk fibroin aqueous solution containing a predetermined amount of the fragrance component was pre-frozen as described above and then freeze-dried, the result was the same as the result of the silk fibroin aqueous solution performed without adding the fragrance component to the sample.

  From the above experiments, for example, to pre-freeze a 0.5% strength silk fibroin aqueous solution at −50 ° C., it is treated for 10 minutes or more, and to freeze-dry at −50 ° C., it is treated for 10 hours or more. It has been found that this is one preferable condition for obtaining solid dry silk containing the fragrance component in the present invention.

  In the following examples, pre-freezing at −50 ° C. was performed for 20 minutes, and freeze-drying at −50 ° C. was performed for 15 hours.

  In the present Example, sensory evaluation of the fragrance with respect to the silk fibroin derived from the rabbit which added rosemary oil was performed.

  GC-MS is an extremely effective analytical means for quantitative determination of fragrance components, but handling requires familiarity with the functions of the apparatus and requires chemical knowledge and skill. Therefore, in this embodiment, instead of using the GC-MS analyzer, the volatilization amount of the scent component of the residual scented silk of the present invention is obtained by sensorically evaluating the scent of the scent component volatilized by a plurality of people from the sample. (The same applies to Examples 3 to 4 and 12 below).

  The experimental method is as follows. Rosemary oil solution (1 drop of rosemary oil stock (0.05 g) was adjusted to 20% with distilled water, respectively, in silk fibroin aqueous solutions derived from rabbits with concentrations of 0.1%, 0.3% and 0.5%. 0.5 g or 1.0 g) was added, and placed in a prefreezing tank (EYELA, PFR-1000) at −50 ° C. for 20 minutes to freeze the silk fibroin aqueous solution. Subsequently, the fragrance component-containing silk fibroin aqueous solution was freeze-dried by treatment for 15 hours using a freeze-dryer (EYELA, FDU-1200) set at −50 ° C. After lyophilization, after a predetermined standing time (24 hours (24 h), 3 weeks (3 w), 6 weeks (6 w)) has elapsed, the sensory evaluation of the fragrance was performed, and the results obtained are shown in Table 1. After the predetermined standing time, water was added to the sample to completely dissolve the solid silk fibroin, and then the sensory evaluation of the aroma was performed again, and the results obtained are shown in Table 2. In Tables 1 and 2, “silk 0.1%, 0.3%, 0.5%” indicates the concentration (wt%) of the used silk fibroin aqueous solution.

  As apparent from Table 1, when silk fibroin containing rosemary oil is left in a dry state, as the standing time (24 hours, 3 weeks, 6 weeks) elapses, even if the silk fibroin concentration changes, Further, even if the concentration of the rosemary oil charged in the initial stage changes, the evaluation value of the rosemary oil aroma due to sensory evaluation is little decreased, and the evaluation value may increase. On the other hand, in the case of the conventional incense component-containing material (the incense component-containing material described in Comparative Examples 1 and 2), most of the incense component from the incense component adsorbing sample is volatilized during the three-week leaving period. have done. Thus, in the case of the conventional fragrance component-containing material, the fragrance is greatly reduced with the volatilization of the fragrance component, but in the case of the fragrance component-containing material of the present invention, there is an effect that the fragrance is substantially maintained or increased. can get.

  On the other hand, in Table 2, after leaving silk fibroin containing rosemary oil in a freeze-dried state for a predetermined time (24 hours, 3 weeks, 6 weeks), a predetermined amount of water (10 mL) is added immediately before the fragrance evaluation. The aroma of each sample was subjected to sensory evaluation after stirring well until the silk fibroin was uniformly dissolved. As a result, in the system to which water was added, the fragrance evaluation value with the passage of the standing time tended to increase as compared with the dry fragrance evaluation value of silk fibroin containing rosemary oil. In addition, when compared with the fragrance evaluation value in the sample in the dry state with the same standing time, the fragrance evaluation value tended to increase in the system to which water was added. In addition, there is no restriction | limiting in particular in the amount of water addition in this case, For example, 3 mL-10 mL are enough effective, and even if it adds the amount of water exceeding 10 mL, it does not affect the result of fragrance evaluation.

  Moreover, in the case of the conventional fragrance component-containing material (fragrance component-containing material described in Comparative Examples 1 and 2), in the system to which water was added, the fragrance evaluation value did not increase and remained slight. Compared to the case of the dried scent component sample of the present invention, the remaining scented property was very weak even when water was added, and the dried scented silk of the present invention did not have the excellent scented property and was poor. It was.

  As a result, the intensity of the aroma of rosemary oil is maintained or increased in the state of being mixed with silk fibroin even if the standing time is increased. However, in the case of a conventional incense component-containing material (the incense component-containing material described in Comparative Examples 1 and 2), if left undisturbed, the aroma decreases greatly as the incense component volatilizes. In contrast, in the case of the incense component-containing material of the present invention, the aroma is maintained as described above. In addition, in the case of the conventional fragrance component-containing material (the fragrance component-containing material described in Comparative Examples 1 and 2), even if water is added, the remaining fragrance property does not become good. Addition suggests that the intensity of fragrance increases (enhances).

(Comparative Example 1)
Put 0.7 g of silk fibroin fiber into the vial used in Example 1 so that it does not touch the fiber directly and non-uniformly (If the scent component touches the fiber directly and non-uniformly, the residual fragrance of the fiber cannot be evaluated correctly. Therefore, 0.1 g of the rosemary oil solution used in Example 2 was added, and the mixture was allowed to stand at room temperature for 4 hours while being sealed. After removing the sample and leaving it to stand for 3 weeks, the residual fragrance of silk fibroin fiber (abbreviated as “fragrance component adsorption sample”) was evaluated by aroma sensory evaluation. It was much lighter than that of the dry silk of the invention, and was almost close to zero. This is because most of the incense components have volatilized during the 3-week standing period. Thus, it was confirmed that the silk fibroin fiber containing the scent component was inferior to the residual scent property of the dry silk containing the scent component of the present invention.

  Subsequently, when water was added to the fragrance component adsorbing sample in the vial, and the odor sensory evaluation was performed again by the method described above, the fragrance evaluation value did not increase and remained slight. Silk fibroin fiber containing a fragrance component does not maintain the remaining fragrance for a long time even if left standing, the remaining fragrance is very weak even when water is added, and there is no excellent remaining fragrance exhibited by the dried silk of the present invention. It was inferior.

(Comparative Example 2)
7% strength rabbit silk fibroin aqueous solution is put in the vial used in Example 1, and 0.5 g of rosemary oil solution is added in the same manner as in Example 2 and placed in a prefreezing apparatus at −50 ° C. for 20 minutes. It was surely frozen. Then, when freeze-dried at -50 ° C. for 15 hours, unlike the dried silk of the present invention in the form of powder, silk fibroin does not become powdery, but is a lump or block of silk with a small surface area. It became fibroin. This is because the concentration of the rabbit silk fibroin aqueous solution used was as high as 7%. According to the odor sensory evaluation, the residual fragrance property obtained by leaving the bulk silk fibroin containing rosemary oil for 3 weeks in a dry state was significantly lower than the residual fragrance property of the dry silk containing the fragrance component of the present invention. The residual fragrance of the bulk silk fibroin was inferior to that of the dried silk of the present invention.

  Subsequently, when water was added to the block / block-like silk fibroin containing the fragrance component and the fragrance sensory test was performed again by the method described above, the fragrance evaluation value did not increase and was extremely weak.

  Thus, the silk fibroin lump containing the fragrance component does not maintain the remaining fragrance property for a long time even if left standing, and the remaining fragrance property is very weak even when water is added. The excellent remaining fragrance exhibited by the dried silk of the present invention Sex could not be confirmed.

  In this example, sensory evaluation of the scent of silk fibroin derived from rabbits with addition of strawberry flavor (natural strawberry juice) was performed.

  0.5 g or 1.0 g of strawberry flavor (natural fruit juice) was added to silk fibroin aqueous solutions having concentrations of 0.1%, 0.3% and 0.5%. Subsequently, in the same manner as in Example 2, preliminary freezing at −50 ° C. for 20 minutes was performed, followed by lyophilization at −50 for 15 hours. After lyophilization, the sample was allowed to stand for a predetermined time (24 hours, 3 weeks), and then the sensory evaluation of the aroma of the sample was performed. The results obtained are shown in Table 3. In Table 3, “silk 0.1%, 0.3% 0.5%” indicates the concentration (wt%) of the silk fibroin aqueous solution used.

  In the above, the evaluation was performed only at 24 hours and 3 weeks as the standing time in order to simplify the experimental conditions.

  As is apparent from Table 3, when the silk fibroin containing strawberry flavor is left in a dry state, the silk fibroin concentration changes as the standing time (24 hours, 3 weeks) elapses. Even if the concentration of the prepared strawberry flavor was changed, the intensity of the fragrance of the strawberry flavor by sensory evaluation tended to be maintained or increased in the case of the fragrance component-containing material of the present invention. On the other hand, in the case of the conventional incense component-containing material (the incense component-containing material of Comparative Examples 1 and 2), as described above, the aroma decreases greatly with the volatilization of the incense component and approaches zero. In addition, even when 10 mL of water was added, the fragrance was not significantly reduced as compared with the conventional fragrance component-containing material and did not become zero.

In this example, sensory evaluation of the scent of silk fibroin derived from rabbit with added vanilla oil was performed. That is, the same fragrance evaluation as in Example 3 was performed using vanilla oil instead of the strawberry flavor used in Example 3. The results obtained are shown in Tables 4 and 5. In Tables 4 and 5, “silk 0.1%, 0.3%, 0.5%” indicates the concentration (wt%) of the used silk fibroin aqueous solution.

  According to Table 4, when the silk fibroin containing vanilla oil is left in a lyophilized state, the concentration of the silk fibroin increases as the standing time (24 hours, 3 weeks) elapses. Even when the oil concentration was increased, the aroma intensity of the vanilla oil by sensory evaluation tended to increase or be maintained. As described above, in the case of the conventional incense component-containing material (the incense component-containing material of Comparative Examples 1 and 2), the aroma is greatly reduced as the incense component evaporates as the standing time elapses. In the case of the scent component-containing material, a special effect of increasing or maintaining the scent was obtained.

  On the other hand, from Table 5, after leaving the silk fibroin containing vanilla oil in a dry state for a predetermined time (24 hours, 3 weeks), 10 mL of water was added and stirred well until the silk fibroin was uniformly dissolved. When the sensory evaluation of the fragrance of each sample, compared with the fragrance evaluation value after leaving the silk fibroin containing vanilla oil in a dry state, the evaluation value is longer when the standing time is longer, There was a tendency to increase or maintain slightly. As described above, in the case of the conventional fragrance component-containing material (the fragrance component-containing material of Comparative Examples 1 and 2), as the standing time elapses, the fragrance decreases greatly with the volatilization of the fragrance component and approaches zero. In the case of the scent component-containing material of the present invention, a special effect that the scent increases or is maintained was obtained.

In this example, the scent component was detected by GC-MS.
A gas chromatograph-mass spectrometer (GC-MS), which is a type of chromatograph, is an analytical instrument that can also be used for identifying and quantifying the fragrance component that is the subject of the present invention. It is characterized in that the sample and mobile phase are gases, and is used as a microanalysis technique in various scientific fields. The equipment used is Shimadzu GCMS-QP5050A.

[Measurement and analysis method]
Conditions for obtaining a stable GC-MS analysis result: An incense component sample was analyzed by flowing helium as a carrier gas at a flow rate of 20 mL / min and a column inlet pressure of 100 kPa. The column oven was heated to 50 ° C., and the temperature of the sample vaporizing chamber and the interface was heated to 50 ° C. In this state, it is preferable to keep operating the apparatus for 6 hours and to sufficiently increase the degree of vacuum of the MS part.

  After confirming that the vacuum was sufficiently high at 30 Pa, the sample vaporization chamber temperature was heated to 250 ° C., the interface temperature was increased to 200 ° C., and analysis of the sample was started. The column oven temperature was increased to 200 ° C. at a rate of 10 ° C./min, the solvent elution time was set to 3 minutes, and the analysis maintenance time was set to 20 minutes. 2 μL of analysis sample was introduced into the apparatus.

[Analytical sample]
GC-MS analysis was performed on the following three fragrance component samples diluted 100-fold with acetone.
(1) Rosemary Oil (Life Wood Co., Ltd., Rosemary Cineol, LOT # 522)
(2) Furaneol (Sigma Aldrich Japan, 2.5-dimethyl-4-hydroxy-3 (2H) -furanone, Lot # STBC7913V)
(3) Vanilla oil (made by Meidi-ya, trade name: Essence vanilla, Lot # 2015.522)

[result of analysis]
Various fragrance components are analyzed by GC-MS, and the obtained GC-MS spectrum (gas chromatogram) is shown in FIGS. FIG. 2 is an analysis result for rosemary oil, FIG. 3 is an analysis result for furaneol, and FIG. 4 is an analysis result for vanilla oil. In each figure, an arrow is attached to the peak having the largest area, that is, the peak corresponding to the component most frequently contained in the sample. The horizontal axis of the gas chromatogram is the retention time (minute), and the vertical axis is the total ion chromatogram (TIC).

The concentration of the incense component was evaluated by applying it to this experiment with reference to the following web page.
https://www.lasoft.co.jp/chromdoc/chrom.asp?sub=chromat6

  As is apparent from the GC-MS spectrum shown in FIG. 2, the main fragrance component contained in rosemary oil is cineole, judging from the retention time of 6.939 minutes corresponding to the peak with the largest area. As can be seen from the GC-MS spectrum shown in FIG. 3, the main fragrance component contained in furaneol is 2,625 minutes, judging from the retention time corresponding to the peak with the largest area of 7.625 minutes. It is 5-dimethyl-4-hydroxy-3 (2H) -furanone and, as is clear from the GC-MS spectrum shown in FIG. 4, the retention time corresponding to the peak with the largest area is 13.575 minutes. Judging from the above, the main flavor component of vanilla oil is vanillin. As is clear from the results of FIGS. 2 to 4, it is possible to identify the fragrance component by examining the retention time of the main peak from the GC-MS spectrum.

  In this example, a rosemary oil, strawberry flavor (furaneol), vanilla oil and rose oil were used, and an analysis experiment of a fragrance component using a column filler was conducted.

  Prior to performing GC-MS quantification of the perfume component sustainedly released from rabbit silk fibroin obtained by pre-freezing and freeze-drying according to the above-mentioned examples carrying these perfume components, optimal experimental conditions A preliminary experiment was conducted to obtain This preliminary experiment was performed using the vaporizer shown in FIG.

  Each of the four types of samples diluted 50-fold with an aqueous ethanol solution adjusted to 50% with distilled water was placed in a 200 mL syringe 1 with 2 mL each in a vial and the lid removed. The syringe inner tube 2 was fixed by being inserted into an outer tube of several centimeters, and a rubber tube was attached to the tip of the syringe and fastened with a pinch cock so that the gas in the syringe would not enter or exit. After standing for 6 hours, 0.1 g of adsorption filler (Grace Davison Discovery Science, trade name: Porapak Q; particle size: 80/100, Batch # 168) is attached to a rubber tube attached to the tip of the syringe. The packed adsorption column 3 was connected to remove the pinch cock, and 150 mL of gas in the syringe was pushed out at a speed of 1 mL / sec to be adsorbed by the adsorbent. FIG. 5 shows a state in which a test gas is introduced into the adsorption column 3 in which the adsorption filler A is enclosed.

  Thereafter, 1 mL of acetone was passed through the adsorbent filler to elute the adsorbed substance, and this solution was introduced into GC-MS. This method is a normal method.

The incense components used were as follows.
(1) Rosemary oil (Life Wooden Co., Ltd., Rosemary Cineol, Lot # 522)
(2) Furaneol (Sigma Aldrich Japan, 2.5-dimethyl-4-hydroxy-3 (2H) -furanone, Lot # STBC7913V)
(3) Vanilla oil (Meidi-ya, Essence vanilla, Lot # 2015.522)
(4) Rose oil (Maggie Tisserand, Essential Oil for Aromatherapy)

[Adsorption onto column packing material]
According to the GC-MS spectrum measurement results, when one drop (0.05 g) of rosemary oil stock solution was diluted 50 times with an ethanol aqueous solution adjusted to 50% with distilled water, the gas chromatogram In addition, a peak almost coincident with the retention time of cineole in Example 5 was observed. However, when one drop (0.05 g) of furaneol and vanilla oil stock solution was diluted 50 times with an ethanol aqueous solution adjusted to 50% with distilled water, no peak corresponding to each major component was seen. It was.

Based on this result, the samples using furaneol and vanilla oil, whose components were not detected, were promoted to vaporize the fragrance components using the following two methods and examined whether they could be detected by GC-MS did.
(1) Normal method (standing for 24 hours): In the normal method described above, the time for standing in a 200 mL syringe was extended from 6 hours to 24 hours.
(2) Heating method: The time for standing in a 200 mL syringe was set to 24 hours, and heating was performed at about 60 ° C. with a drier from the outside of the syringe for the first hour.

  However, these methods still failed to detect the components in the samples using furaneol and vanilla oil.

Therefore, detection by GC-MS was attempted using rose oil (Maggie Tisserand, Essential Oil for Aroma-therapy) as a new scent component. The component contained in the sample was not detected by the normal method, but the component was detected by the heating method although the detection intensity was very small.
The above results are shown in Table 6 below.

In order to examine whether detection is possible by GC-MS measurement, the evaluation described in Table 6 has the following meaning.
○: Indicates that detection is possible.
(Triangle | delta): Although detection is possible, detection intensity is very small.
X: Detection is not possible.
-: No experiment was conducted.

  From the above results, cineol and phenethyl alcohol may be easier to detect by GC-MS than furaneol or vanillin. Therefore, in Example 7 below, a sample was prepared using rosemary oil and rose oil as a scent component.

In this example, GC-MS analysis was performed by a normal method using a column filler and a heating method.
The following incense ingredients were used to examine the conventional method and heating method.
(1) Rosemary oil (Tree of Life, Rosemary Cineol, Lot # 522)
(2) Rose oil (Maggie Tisserand, Essential Oil for Aromatherapy)

[Preparation of sample for GC-MS measurement]
A mixture of ethanol and distilled water in a ratio of 1: 1 (50% ethanol) was used as a diluent. First, 2 mL of the prepared silk fibroin aqueous solution having a concentration of 0.5% or 1.5% was dropped into the vial. One drop (0.05 g) of the fragrance ingredient stock solution was diluted 50-fold with a diluent, 1 mL was added to each silk fibroin aqueous solution, pre-frozen at -50 ° C for 20 minutes, and then lyophilized at -50 ° C for 15 hours.

[Measurement conditions for GC-MS]
The sample that was left for a predetermined time after removing the lid of the vial was placed in a 200 mL syringe together with the vial and allowed to stand for 6 hours. The gas was extruded at a rate of 1 mL / sec, 150 mL, and passed through a packed column. 1 mL of acetone was poured into the column, and the liquid that emerged was collected. Each component was quantified using GC-MS. In addition, after analyzing the sample in the dry state, water was added to the sample, and gas was collected and analyzed in the same manner.

  Pre-frozen and freeze-dried silk fibroin containing various fragrance components is placed in the vaporizer shown in FIG. 5 described in Example 6 and allowed to stand for 6 to 24 hours, after which it is adsorbed onto an adsorption filler (Porapak Q). Then, the acetone eluate eluted with acetone was introduced into GC-MS, and it was examined whether the main compound of the scent component could be detected with GC-MS. This is the normal method.

  If the normal method fails to detect, the pre-frozen and freeze-dried silk fibroin containing the fragrance component is put into a vaporizer, and the internal temperature of the device is heated to about 60 ° C with a dryer for 1 hour from the outside of the vaporizer. did. After standing for 24 hours including heating time, the fragrance component to be vaporized is passed through an adsorption column packed with an adsorption packing material (Porapak Q) and then adsorbed by flowing acetone through the adsorption packing material. It was. It was examined whether it could be detected by GC-MS by introducing it into GC-MS. This is a heating method.

  Table 7 shows the results of GC-MS analysis performed by a conventional method or a heating method using rosemary oil and rose oil as the fragrance component.

In order to examine whether detection is possible by GC-MS measurement (normal method), the evaluation described in Table 7 was performed as follows.
○: Indicates that detection is possible.
X: Detection is not possible.
-: No experiment.

  In the case of using pre-frozen and freeze-dried silk fibroin containing fragrance components of rosemary oil and rose oil, neither the conventional method nor the heating method could be evaluated by GC-MS.

  In this example, an experiment was conducted in which a fragrance component was directly introduced into GC-MS without using a column filler.

  There may be a problem in the combination of the column filler or the fragrance component once adsorbed on the column filler and the elution solvent for flowing out from the column filler. Therefore, it was next decided to introduce the gas in the upper space of the vial directly into the GC-MS without using a column filler.

  The fragrance component to be used and the method for preparing the sample were the same as in Example 7.

[Measurement conditions for GC-MS]
The vial lid was removed and the sample left for a predetermined time was allowed to stand for 6 hours with the vial lid closed. Unlike Example 7, without using an adsorption column containing a column filler, an injection needle was directly placed in the rubber lid of a glass vial, and 5 mL of gas was collected from the upper space (head space) of the bottle. This gas was extruded at 1 mL / second using a syringe and introduced into the GC-MS.

Rosemary oil could be analyzed by GC-MS. The area of the peak showing cineol as the main component was 6.79 × 10 ⁶ . In rose oil, a small peak corresponding to phenethyl alcohol was detected.

  Pre-frozen and lyophilized silk containing rosemary oil solution sealed in a vial was left at room temperature for 3 days after the experiment, with the rubber lid of the vial completely removed. After standing, the vial was attached to the vial with a rubber lid and allowed to stand for 6 hours, and then an injection needle was inserted into the rubber lid of the vial, and 5 mL of gas was collected from the upper gap of the bottle into the syringe. The collected gas was extruded from the syringe at a rate of 1 mL / second and introduced into the GC-MS.

  According to the GC-MS measurement results, the main peak appearing in the gas chromatogram was due to cineole from the corresponding retention time.

  Furthermore, after completely removing the rubber lid of the vial containing the pre-frozen and freeze-dried silk fibroin containing the rosemary oil solution and allowing to stand at room temperature for 3 days, 2 mL of distilled water was added to the vial. The solid silk fibroin was thoroughly stirred until it was completely dissolved, and was allowed to stand for 6 hours in a sealed state with a rubber lid attached. The injection needle was directly placed in the rubber lid of the vial, and 5 mL of gas was collected from the upper gap of the bottle with a syringe. This gas was extruded at 1 mL / second using a syringe and introduced into the GC-MS.

According to the GC-MS measurement results, the main peak appearing in the gas chromatogram was due to the cineole peak from the corresponding retention time. A vial containing pre-frozen and lyophilized silk containing a rosemary oil solution, compared to the case where the rubber lid was removed and the sample was not left in the atmosphere or was left for 3 days and analyzed without adding distilled water. After completely removing the rubber cap from the bottle and allowing to stand at room temperature for 3 days, add 2 mL of distilled water to the vial and stir well until the solid silk is completely dissolved. When the gas in the vial was collected and analyzed for 6 hours, the peak area corresponding to cineol increased. The peak area was 20.22 × 10 ⁶ .

  It was confirmed that the area of the peak corresponding to cineol contained in the gas in the vial was increased by adding water.

  Table 8 below relates to the peak corresponding to the main component cineol when analyzed by GC-MS of the gas in the upper void in the vial containing the pre-frozen and lyophilized silk sample containing the rosemary oil solution. It is data.

  Table 8 shows the results of GC-MS analysis of the gas before and after opening the bottle lid.

Samples and symbols in Table 8 mean the following.
“Add water after opening” means a gas collected from the upper space of a bottle in which water is added to silk fibroin in the bottle after the rubber stopper is opened for 3 days;
T1: peak retention time in GC-MS analysis (minutes);
T2: Peak start time (in minutes) in GC-MS analysis;
T3: Peak end time (in minutes) in GC-MS analysis;
Area: peak area;
Height: Peak height.

  Remove the rubber lid of the vial containing the pre-frozen and freeze-dried silk fibroin sample containing the rosemary oil solution and keep it open for 3 days, and then pre-freeze and freeze-dry the solution containing the rosemary oil solution. When distilled water is added to a silk fibroin sample, the peak area of the gas chromatogram corresponding to the slowly released fragrance component increases, and the fragrance component that is gradually released from the freeze-dried silk fibroin by adding water may increase rapidly. confirmed.

  This indicates that GC-MS was able to confirm the correctness of the results of the examples obtained by the sensory test on a sample obtained by adding water to a pre-frozen and freeze-dried silk fibroin sample containing a rosemary oil solution. This means that the inventive step of the scented silk of the present invention is demonstrated.

  In this example, the quantification by GC-MS of the scent component released from dried silk derived from rabbits was examined.

  Incense component used: Rosemary oil (cineole) (Life Wood Co., Ltd., Lot # 522). As a diluted solution of this fragrance component, a mixture of ethanol and distilled water at a ratio of 1: 1 (50% aqueous ethanol solution) was used.

  For the sample, add 2 mL of a silkworm-derived silk aqueous solution prepared to a concentration of 0.5% or 1.5% to the vial, dilute one drop (0.05 g) of the fragrance ingredient stock 50 times with the diluent, The diluted scent component solution was prepared by adding 1 mL each to the silk aqueous solution. This aqueous solution was pre-frozen at -50 ° C for 20 minutes, followed by lyophilization at -50 ° C for 15 hours.

[experimental method]
(1) The rubber stopper of the vial was opened and left for a predetermined time (24h, 1w, 3w, 6w).
(2) The rubber stopper was closed and allowed to stand for 6 hours.
(3) The needle tip of the syringe was inserted into the vial, and 5 mL of gas above the sample was collected.
(4) The collected gas was introduced into GC-MS to quantify each scent component.
After analysis of the dried sample, water was added to make it wet, and gas was collected and analyzed in the same manner.

[Experimental result]
GC-MS analysis was performed using samples containing rosemary oil (cineole) at different standing times, and the peak area of cineole was determined. The results obtained are summarized in Table 9.

  In Table 9, “Cineol” means “cineole”, “Silk 0.5%” means “0.5% silk aqueous solution”, and “Silk 1.5%” means “1.5% silk aqueous solution”. "" Indicates no detection, and "dry" and "wet" indicate a dry sample and a wet sample with added water, respectively.

  The results of the sustained release of cineole from rosemary oil-containing dry silk shown in Table 9 (comparison in terms of peak area) are shown in FIG.

In FIG. 6, the symbols are as follows.
○: Silk 0.5%, dry ●: Silk 0.5%, wet □: Silk 1.5%, dry ■: Silk 1.5%, wet

  The following became clear from Table 9 and FIG. When the sustained release amount of cineole from freeze-dried silk containing rosemary oil was compared with the peak area by GC-MS measurement, it was obtained from 0.5% and 1.5% silk aqueous solution in the dry state after standing for 3 weeks. In the obtained freeze-dried silk, the residual scent of cineole is almost maintained, and it is effective compared to the large decrease in the case of the conventional scent component-containing materials (scent component-containing materials of Comparative Examples 1 and 2). To be judged. Moreover, when water was added to freeze-dried silk obtained from 0.5% and 1.5% silk aqueous solution containing rosemary oil, the sustained release amount of cineol from the wet sample became a large value. . That is, it was confirmed that when water was added to freeze-dried silk obtained from 0.5% and 1.5% silk aqueous solutions containing cineole, the sustained release amount of cineole increased specifically. Thus, even when water is added, the residual fragrance of cineol is almost maintained, and the effect is larger than the large decrease in the case of the conventional fragrance component-containing material (the fragrance component-containing material of Comparative Examples 1 and 2). It is judged that there is.

  In this example, the sustained release amount of cineol from silk fibroin derived from rabbits in a pre-frozen / lyophilized state was measured.

  Add 1 mL of a solution obtained by diluting cineole 50 times with 2 mL of 0.5%, 1.0%, 1.5% silk fibroin aqueous solution by mixing ethanol and distilled water at a volume ratio of 50:50. Unlike the case of Example 7, this was packed in a vial with a rubber stopper without using an adsorption column containing a column filler, pre-frozen at −50 ° C. for 20 minutes, and then at −50 ° C. Lyophilized for 15 hours. Next, the rubber stopper of the vial is opened and left for a predetermined time (24 hours, 1 week, 3 weeks, 6 weeks), and then the vial is closed for 6 hours with the rubber stopper closed. A syringe needle was attached and inserted into the vial, and 5 mL of gas on the powder sample, that is, in the upper space of the vial was collected, and components contained in this gas were quantified by GC-MS. After analyzing the dried sample in this way, subsequently, after a predetermined period of time, 2 mL of water is added to the dried sample, and a syringe needle is attached and inserted into a vial. 5 mL of gas in the upper gap of the vial was collected to quantify the gas component.

  As a result, in the experiment of 0.5% silk fibroin aqueous solution, when the sample was in a dry state, the amount of cineole sustainedly released from the sample was already very small immediately after opening according to the peak area in the GC-MS measurement. there were. When left for 6 weeks, the sustained release amount decreased as the detection by GC-MS was more difficult. When water was added to the dried sample, the amount of cineole that was gradually released from the sample was slightly increased. However, according to the peak area in the GC-MS measurement, when the concentration of the silk fibroin aqueous solution is 1.5%, the sustained release amount of cineol from the sample in the dry state is the result of the above 0.5% silk fibroin aqueous solution. However, when water was added, the sustained release amount of cineol from the sample was a value about several tens of times greater than the sustained release amount of cineol from the dried sample. This is because the amount of sustained release of cineol from the dry sample is very small, but when the silk fibroin aqueous solution concentration is 1% or more, when water is added, the cineol in contact with the silk is separated from the sample and released in a large amount. Is meant to do. In addition, when a 1.0% silk fibroin aqueous solution was used, the same results were obtained as is clear from the results shown in Table 10. Furthermore, it was found that the above results were almost maintained even when left for a predetermined time, and that there was a residual scent. In addition, a favorable result can be obtained when the silk fibroin aqueous solution density | concentration is 0.4 to 4 weight%.

  In this example, a residual fragrance experiment using silk fibroin derived from silkworms was performed. Instead of silk fibroin derived from rabbits used in Example 9, a 0.5% silk fibroin aqueous solution of wild silkworms was used, and 1 drop (about 0.05 g) of rosemary oil was distilled 50% with distilled water. The mixture prepared by adding 50-fold diluted ethanol aqueous solution was put into a glass vial with a rubber stopper, pre-frozen at -50 ° C for 20 minutes, and then lyophilized at -50 ° C for 15 minutes. Let dry for hours. Without using an adsorption column containing a column filler, an injection needle was directly placed in the rubber lid of a glass vial, and 5 mL of gas was collected from the upper space of the bottle. According to the analysis by GC-MS, the peak area indicating cineol, which is the main component of rosemary oil contained in the gas in the vial, was larger than that in Example 10. It was revealed that the sustained release amount of cineole from cocoon silk fibroin containing cineole is larger than that of rabbit silk fibroin. From this, in the case of silk fibroin derived from silkworms, if the above evaluation is performed on a silk fibroin derived from silkworms, the peak area increases more than in the case of silk fibroin derived from silkworms. It is considered a thing. Furthermore, it was found that the above results were almost maintained even when left for a predetermined time, and that there was a residual scent.

  From the above results, it was found that the silk fibroin derived from wild silkworm has higher residual fragrance function for the scent component than silk fibroin derived from rabbit. This seems to be related to the structural properties of silk fibroin from wild silkworms. That is, the silk fibroin crystal structure derived from wild silk is composed of poly-L-alanine composed of alanine chains, and is characterized by a dense structure in which intermolecular and intramolecular hydrogen bonds are firmly taken. For this reason, the thermal decomposition temperature of silk fibers derived from wild silkworms is 355 ° C, whereas the thermal decomposition temperature of silk fibers derived from rabbits is different from 310 ° C. When the scent component binds to the wild silk fibroin having a dense structure with a strong interaction, the residual scent of the wild silk fibroy containing the scent component is excellent. The main molecular structure of wild silk fibroin powder or film-like sample is a random coil and α helix, and the conversion to β structure occurs clearly by alcohol treatment. It is considered that fibroin can be preferably used as a carrier for aroma component molecules.

  In this example, a residual fragrance experiment was performed using a mixed material of silk fibroin derived from rabbits and wild silkworms.

  In place of the silk fibroin aqueous solution derived from rabbits used in Example 9, a silk fibroin aqueous solution derived from rabbits having the same concentration of 0.5 wt% and 1 wt%, respectively, and a silk fibroin aqueous solution derived from wild silkworms had a volume ratio of 50:50. 1 ml of rosemary oil solution (1 drop (about 0.05 g) of rosemary oil stock solution) was distilled into a mixed aqueous solution (two types of mixed aqueous solution) of rabbit silk fibroin and wild silk fibroin combined with (Mixed 50 times with an aqueous ethanol solution adjusted to 50% with water) was added to a glass vial with a rubber stopper, pre-frozen at -50 ° C for 20 minutes, and then frozen at -50 ° C. It was made to dry with a dryer for 15 hours. Without using the glass tube of the adsorption column containing the column filler, an injection needle was directly placed in the rubber lid of a glass vial, and 5 mL of gas was collected from the upper space of the bottle. According to the analysis by GC-MS, the peak areas indicating cineol, which is the main component of rosemary oil contained in the gas in the vial, were both larger than those in Example 10. It became clear that the sustained release amount of cineole from the mixed material of silk fibroin with rabbits and wild silk containing cineole increased more than that of rabbit alone or wild rabbit alone. The same can be said when water is added. Furthermore, it was found that the above results were almost maintained even when left for a predetermined time, and that there was a residual scent.

  10 mL of an aqueous silk solution prepared by adding rosemary oil (cineole) as a fragrance component to a 0.5 wt% silk aqueous solution derived from rabbits was prepared. In this case, according to Example 10, 1 mL of a mixture of ethanol and distilled water in a ratio of 1: 1 (50% aqueous ethanol solution) was used as a diluted solution of the fragrance component. The silk aqueous solution containing the fragrance component thus prepared was spread on the surface of the substrate (polystyrene plate), and the aqueous solution was evaporated and dried at room temperature for a whole day and night to prepare a transparent silk film containing the fragrance component. Furthermore, dilute acetic acid is added to the silk aqueous solution thus prepared, the pH is adjusted to 3.5, the mixture is allowed to stand for 6 hours, pre-frozen at −50 ° C. for 20 minutes, and then freeze-dried at −50 ° C. for 15 hours. A sponge-like silk porous body containing a scent component was produced.

  Immediately after a predetermined time (24 hours, 3 weeks, 6 weeks) has elapsed in a standard state of 10% RH at 25 ° C., each of the transparent silk film and silk porous body containing the scent component is placed in a vial. A human evaluator performed a sensory evaluation of the fragrance in the Bayer bottle in accordance with a 4-step sensory odor evaluation standard. After the predetermined standing time had elapsed, 10 mL of distilled water was added to the sample, and after stirring well, sensory evaluation of the aroma was performed again.

  In the cineol-containing silk membrane and the silk porous body, the evaluation value of the fragrance is hardly lowered even when the standing time has elapsed, and when 10 mL of distilled water is added, the evaluation of the fragrance is compared with that before adding water. It was observed that the value increased. Furthermore, it was found that the above results were almost maintained even when left for a predetermined time, and that there was a residual scent.

  From these results, it was found that, in addition to powdered silk, silk membranes and silk porous bodies have a residual fragrance as well.

  A silk aqueous solution prepared by adding 1 mL of rosemary oil stock solution (0.05 g) diluted 50 times with an aqueous ethanol solution adjusted to 50% with distilled water to a silk solution derived from a rabbit prepared to a concentration of 1.0%. A silk solution derived from rabbits having a concentration of 1.0% and containing no rosemary oil was placed in a prefreezing tank (EYELA, PFR-1000) at −50 ° C. for 20 minutes to freeze the silk solution. Subsequently, a dry silk containing a scent component or a dry silk containing no scent component was produced by performing a drying treatment for 15 hours using a freeze dryer (EYELA, FDU-1200) set to −50 ° C. In the above-described preliminary freeze-drying and freeze-drying, vials were used. In addition, in the case of the silk aqueous solution which does not contain a fragrance | flavor component at all, what diluted the said rosemary oil was added with respect to the manufactured dry silk.

  After the silk in the dried state was obtained, the quantitative determination of the fragrance components in the vials was evaluated by GC-MS measurement after a predetermined standing time, according to the method of the above example. Furthermore, after the predetermined standing time has elapsed, 10 mL of distilled water is added to the sample in the vial to completely dissolve the solid silk fibroin, and then the fragrance component in the vial is quantified according to the method of the above example. It evaluated by GC-MS measurement. The results obtained are summarized in Table 10.

  In Table 10, “-” indicates that cineole could not be detected. Further, Silk 1.0% means a silk aqueous solution having a concentration of 1.0%.

  From Table 10, when the fragrance component was added before lyophilization, the following was found by the sustained release experiment of rosemary oil using silk solution derived from rabbits. When silk from rabbits is used as a sustained release carrier, the addition of water to dry silk containing cineol further increases the sustained release of cineol from the sample, and the fragrance after standing for 3 weeks. The fragrance of the component was almost maintained without significant reduction. In addition, the sustained release amount of cineol in the standing time of 24 hours for silk derived from rabbits containing cineol in the dry state is the peak area ratio compared to hydrolyzed silk in the dry state containing cineol carried out in the same manner as described above. The former is about 4 times larger, and the sustained release amount of cineol from silk derived from rabbits containing cineol with 10 mL of water added after 24 hours of standing includes cineole with 10 mL of water added. Compared to hydrolyzed silk, it was 1.6 times larger.

  Further, from Table 10, when the fragrance component was added after lyophilization, that is, after adding a predetermined amount of the fragrance component to lyophilized silk not containing cineol, it was allowed to stand for 24 hours and subjected to GC-MS measurement. The peak area was large, and a large amount of sustained release of cineole was observed. Furthermore, it was found that when a predetermined amount of fragrance component was added to freeze-dried silk containing no cineole and then allowed to stand for 3 weeks, a large amount of cineol was gradually released from the sample, and the residual amount of cineole was greatly reduced. The same tendency was observed when 10 mL of water was added.

  Thus, as is apparent from Table 10, the scent component should be added to the silk aqueous solution before lyophilization, and when added after lyophilization, the sustained release of cineol becomes noticeable as the standing time elapses, and the residual amount decreases. To do.

Instead of silk derived from rabbits used in the above examples, polyvinyl alcohol (PVA) having a polymerization degree of 2000 and a saponification degree of 99.1 mol% was used to determine the sustained release amount of cineol from a sample containing cineol. It was evaluated by measurement. From the comparison of the results of GC-MS measurement, the peak areas of cineole in silk from rabbit and PVA are 83.74 × 10 ⁶ and 51.62 × 10 ⁶ respectively, and the silk from silk that is excellent in rabbit is excellent. It was found to be a release substrate.

In this example, the hydrophilic and hydrophobic characteristics of silk fibroin membranes derived from rabbits were examined.
30 mg of silk derived from rabbits and 30 mg of hydrolyzed silk (manufactured by Ichimaru Falcos), which was used as a comparative sample, were separately put in 2 mL of distilled water to completely dissolve the silk. The film was spread on the surface of a 0.2 mm thick polypropylene film (trade name: Clear Folder A4, manufactured by Cainz Co., Ltd.).

  In addition, when the color change of the aqueous solution obtained by dissolving silk from silkworms and hydrolyzed silk in distilled water was observed, all samples were found when silk from silkworms was dissolved in distilled water. It quickly dissolved and the aqueous solution became slightly translucent. When hydrolyzed silk was dissolved in distilled water, all samples were dissolved neatly, and the aqueous solution became transparent, but turned slightly yellow due to the slightly colored powder pigment.

  Rabbit-derived silk and hydrolyzed silk are each dissolved in distilled water, allowed to stand at room temperature for 3 days, and the moisture in the sample is gradually evaporated to form a solid silk film on the polypropylene film. Could be manufactured. The shape of the obtained silk fibroin membrane was examined.

  In the case of silk derived from rabbits, a circular thin film having a diameter of 5 cm and a thickness of 20 μm was obtained, which was a transparent silk thin film and was transparent and rich in flexibility.

  On the other hand, when hydrolyzed silk was used, it became a circular silk film, and its diameter was 8 mm (thickness 1 mm). Silk solids obtained from hydrolyzed silk are transparent but brittle and the film strength is lower than that of silk derived from rabbits.

  From the above, it was suggested that the molecular weight of silk derived from rabbits is considerably high, and that hydrolyzed silk is low molecular weight. Therefore, in the case of dry silk obtained from silk fibroin derived from rabbit having a high molecular weight, the residual fragrance is higher than in the case of hydrolyzed silk.

  In this example, the gelation of a silk aqueous solution (silk fibroin aqueous solution derived from rabbits) was examined.

  The same silkworm-derived silk and hydrolyzed silk as in the above examples were each dissolved in 5 mL of distilled water and completely dissolved. When the solution was allowed to stand at room temperature for 2 days, the aqueous solution in which silkworm-derived silk was dissolved completely gelated (hardened), but hydrolyzed silk remained in an aqueous solution state even after 16 days from dissolution, and the aqueous solution became opaque. There was no gelation. It was suggested that silk derived from rabbits has a high molecular weight.

  According to the present invention, it is possible to provide a perfume silk that contains a perfume component and has excellent perfume sustained release, perfume persistence, and washing durability, and a method for producing the same, and a simple perfume component detection method. It can be used in the food field, clothing field, interior field, bathing agent, and other industrial fields.

1 Syringe 2 Syringe inner tube 3 Adsorption column A Adsorption filler

Claims (7)

Rosemary oil, rose oil, and the fragrance component selected from the strawberry flavor, at a temperature of the silk solution silk Fiburoi emissions from silkworm -10 ℃ ~-80 ℃, and time pre-freezing of 9 minutes to 60 minutes And then freeze-dried at −50 ° C. so as to completely sublimate the moisture contained in the flavor component and the silk fibroin derived from the rabbit that has been frozen and solidified by this preliminary freezing. It is, that the production of substantivity silk consisting of solid dry silk and a silk fibroin derived flavoring components and the silkworm, and substantivity silk produced is the addition of water to be used at the time of use A method for producing a scented silk characterized by the following. The silkworm derived drying silk, the production method of substantivity silk according to claim 1, characterized in that the molecular structure is have a structure of shaped random coil. An incense ingredient selected from rosemary oil, rose oil and strawberry flavor, silk fibroin derived from wild silkworm, and silk fibroin silk aqueous solution selected from a complex of silk fibroin derived from rabbit and silk fibroin derived from wild silkworm Is pre-frozen at a temperature of −10 ° C. to −80 ° C. for a period of 9 minutes to 60 minutes, and then the incense component frozen and solidified by this preliminary freezing and the silk fibroin derived from the wild silkworm or from the wild silkworm Freeze-dried at −50 ° C. to completely sublimate the water contained in silk fibroin and the silk fibroin complex derived from the rabbit, and the fragrance component and the silk fibroin derived from the wild silkworm, or the fragrance component Fragrance comprising a solid silkworm-derived dried silk comprising a silk fibroin derived from the silkworm and a silk fibroin complex derived from the silkworm To produce silk, or to produce a scented silk made of a composite of this solid wild silk-derived dried silk and solid rabbit-derived dried silk. method for producing a substantivity silk characterized by Rukoto be used by adding water. Drying silk from the silkworm is the molecular structure has the structure of a form random coil, drying silk from the wild silkworm, characterized in Rukoto the molecular structure having a molecular structure of the helix and random coil α The manufacturing method of the aftercense silk of Claim 3 . The freeze-drying, at -50 ° C., 3 hours is performed 15 hours substantivity silk method according to any one of claims 1 to 4, characterized in Rukoto. Method for producing a substantivity silk according to any one of claims 1 to 5, the concentration of the silk solution of the silk fibroin, characterized in that Ru 0.5 to 1.5 wt% der. Solid dry silk obtained After the freeze drying, substantivity according to any one of claims 1 to 6, wherein the dry silk der Rukoto powdered dry silk or porous body Method for producing silk.

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