JP6423998B2 - Supercooling accelerator and method for producing supercooling accelerator - Google Patents

Description

The present invention is, supercooling promoting agent, and relates to a manufacturing method of the supercooling promoting agent.

  The phenomenon that the liquid does not solidify even when the temperature is lower than the melting point is generally called a supercooling phenomenon.

  Conventionally, various supercooling accelerators (anti-ice nucleation activators) for promoting such a supercooling phenomenon are known, for example, those containing an extract extracted from straw. (Patent Document 1).

  By adding such a supercooling accelerator to a liquid containing water, for example, the lower limit of the temperature at which the supercooling phenomenon occurs in the liquid after the addition can be made lower than before the addition.

  However, although such a supercooling accelerator has a supercooling promoting ability, it has a problem that the antioxidant performance is not always sufficient. That is, such a supercooling accelerator has a problem that it does not necessarily have both a supercooling promoting ability and an antioxidant performance.

JP 2010-121052 A

In view of the above-described problems, an object of the present invention is to provide a supercooling accelerator that has both supercooling promoting ability and antioxidant performance.
Moreover, this invention makes it a subject to provide the manufacturing method of the supercooling promoter which can obtain the supercooling promoter which has fully supercooling promotion capability and antioxidant performance .

The supercooling accelerator of the present invention contains a compound represented by a composition formula of C ₁₅ H ₂₆ N ₂ O ₇ extracted from coffee beans, and the compound contains at least an aromatic hydrocarbon structure, a carboxy group in the molecule. It is characterized by having.

  In the supercooling accelerator of the present invention, the coffee beans are preferably roasted coffee beans.

The method of manufacturing supercooling promoting agent of the present invention, at least the aromatic hydrocarbon structure and possess a carboxyl group C ₁₅ H ₂₆ N supercooling promoting agent comprising a compound represented by the composition formula ₂ O ₇ in a molecule A manufacturing method of
A water extraction step of obtaining a water extract by subjecting the coffee beans to extraction with water;
A fractionation step of obtaining a fraction solution containing the compound by subjecting the water extract extracted by the water extraction step to a fractionation treatment according to the molecular weight;
A pH adjusting step for adjusting the pH of the fractionated liquid containing the compound to the acidic side;
and an organic solvent extraction step of performing an extraction treatment with an organic solvent on the pH-adjusted fraction solution .

The supercooling accelerator of the present invention has the effect of sufficiently combining supercooling promoting ability and antioxidant performance. In addition, the method for producing a supercooling accelerator of the present invention produces an effect that a supercooling accelerator having sufficient supercooling promoting ability and antioxidant performance can be obtained .

The chart which shows the result of anion exchange chromatography. The chart which shows the result of a gel filtration chromatography. The figure showing the LC-MS analysis result of a compound. The figure showing the MS-MS analysis result of a compound. Diagram illustrating The ¹ HNMR analysis of Compound. The figure showing the evaluation result of supercooling promotion ability. The figure showing the evaluation result of supercooling promotion ability. The figure showing the evaluation result of antioxidant performance.

  Hereinafter, an embodiment of the supercooling accelerator according to the present invention will be described in detail.

  The supercooling accelerator of this embodiment contains a compound extracted from coffee beans, and the compound has at least an aromatic hydrocarbon structure and a carboxy group in the molecule.

  The coffee beans are seeds of plants belonging to the genus Coffea.

  Examples of plants belonging to the genus Coffea include arabica species (Coffea arabica) and robusta species (Coffea canephora).

The coffee beans are preferably roasted in that the supercooling promotion performance and the antioxidant performance are more excellent.
The roasting is heating coffee beans at a temperature exceeding the boiling point of water. As temperature in roasting, 150-200 degreeC is preferable.

The extraction is to subject the coffee beans to an extraction process with an extraction solvent to obtain at least an extract after the extraction process.
Details of the extraction process will be described later.

The compound has at least an aromatic hydrocarbon structure and a carboxy group in the molecule.
For example, the compound is represented by a composition formula of C ₁₅ H ₂₆ N ₂ O ₇ and has a molecular weight of 346 when the carboxy group is not a salt.

Although the content rate of the compound in the said supercooling promoter is not specifically limited, For example, it is 10-100 mass%.
In addition, although the said supercooling promoter is a liquid aspect normally, it can also be a powder form and a paste-form aspect.

  Next, an embodiment of a method for producing a supercooling accelerator according to the present invention will be described in detail.

The method for producing a supercooling accelerator of this embodiment is a method for producing a supercooling accelerator containing a compound having at least an aromatic hydrocarbon structure and a carboxy group in the molecule,
The compound is extracted by subjecting a water extract extracted from coffee beans with water to an extraction treatment with an organic solvent.

  Preferably, in the method for producing a supercooling accelerator of the present embodiment, a water extraction step of obtaining an aqueous extract by subjecting coffee beans to extraction with water, and a pH adjustment step of adjusting the pH of the water extract And an organic solvent extraction step of obtaining an extract from which the above compound has been extracted by subjecting the pH-adjusted water extract to an extraction treatment with an organic solvent. According to such a production method, there is an advantage that the concentration of the compound in the extract containing the compound described above can be increased efficiently.

In the manufacturing method of the said supercooling promoter, the roasting process which roasts coffee beans can be performed before a water extraction process.
In the method for producing a supercooling accelerator, after the water extraction step and before the pH adjustment step, a fractionation treatment according to the molecular weight is performed on the water extract after the water extraction treatment containing the compound described above. The fractionation process which performs can be performed.
In the manufacturing method of the said supercooling promoter, the refinement | purification process which performs a refinement | purification process to the extract after the extraction process containing the said compound can be performed after an organic-solvent extraction process.

That is, the method for producing the supercooling accelerator is:
Roasting process to roast coffee beans;
A water extraction step of subjecting the roasted coffee beans to an extraction process with water;
A fractionation step of obtaining a fraction solution containing the above compound by subjecting the water extract extracted by the water extraction step to a fractionation treatment according to the molecular weight;
A pH adjustment step of adjusting the pH of the fractionated solution containing the above compound;
an organic solvent extraction step of performing extraction with an organic solvent on the pH-adjusted fraction,
And a purification step of subjecting the extract after the extraction treatment with an organic solvent to a purification treatment.

  In addition, before the water extraction step, the coffee beans can be more easily extracted by crushing the coffee beans.

In the roasting step, the above-described coffee beans are roasted. In the roasting process, as described above, a temperature exceeding the boiling point of water is added to the coffee beans.
In the roasting process, a general method of roasting coffee beans is employed.
In the roasting step, the roasting temperature is preferably 150 to 200 ° C.

In the water extraction step, the coffee beans are extracted with a water-containing extraction solvent containing water by a general extraction process. And the water extract extracted with the water-containing extraction solvent is obtained. The aqueous extract contains the above compound.
Examples of the coffee beans subjected to the extraction treatment include roasted coffee beans or a residue (coffee mash) obtained by once extracting the roasted coffee beans with water.

In the water extraction step, water is usually employed as the water-containing extraction solvent. In addition, as a water containing extraction solvent, what contains 90 mass% or more of water is employable.
In the water extraction step, it is preferable to adjust the pH of the water-containing extraction solvent to 6-8.

In the said water extraction process, extraction temperature is 90-100 degreeC normally. The extraction time is, for example, 30 minutes to 24 hours.
The water extraction step may be performed in a pressurized state by using, for example, an autoclave.

In the water extraction step, for example, the volume ratio of the water-containing extraction solvent to 1 part by mass of dried coffee beans is 1 to 5 times.
In the water extraction step, operations such as filtration, precipitation, centrifugation, and drying are appropriately performed as necessary.
The water extract obtained by the water extraction step contains the compound described above. The water extract can be in the form of liquid or powder.

  In the water extraction step, the water extraction process may be performed a plurality of times. For example, in the water extraction step, a residue after the coffee beans are extracted with water may be obtained, and the residue may be further extracted with water to obtain a water extract.

In the fractionation step, components contained in the water extract obtained in the water extraction step are fractionated according to the molecular weight by a general fractionation method.
As a fractionation treatment method, a fractionation treatment method using a dialysis membrane capable of fractionation with a predetermined molecular weight, a fractionation treatment method using ion exchange chromatography, a fractionation treatment method using an ultrafiltration membrane, and the like can be employed.

  For example, in the fractionation step, a fractionation treatment solution is prepared by, for example, diluting a water extract with water. And the component exceeding predetermined molecular weight and the component below predetermined molecular weight are obtained using the ultrafiltration membrane by which the molecular weight cut off was set to the predetermined value.

  Moreover, in the said fractionation process, the component contained in the water extract obtained by the water extraction process is fractionated into the component with a molecular weight exceeding 10,000 and the component with a molecular weight of 10,000 or less, for example. And the fractionated liquid containing the component of molecular weight 10,000 or less fractionated will contain said compound, and will be used by the subsequent pH adjustment process.

  In the pH adjustment step, the pH of the fractionated fraction is adjusted by a general method. In the pH adjusting step, for example, the pH of the fractionated liquid containing fractionated components having a molecular weight of 10,000 or less is adjusted.

In the pH adjusting step, it is preferable to adjust the pH of the fractionated fraction solution to the acidic side in that the carboxyl group of the compound described above is prevented from becoming a salt form.
By adjusting the pH to the acidic side, the water solubility of the above-described compound becomes smaller, and there is an advantage that the above-mentioned compound can be more reliably extracted in the subsequent organic solvent extraction step.

In the pH adjustment step, for example, an inorganic acid such as hydrochloric acid or sulfuric acid or an organic acid such as acetic acid or citric acid can be added to the fraction solution in order to adjust the pH to the acidic side.
Specifically, in the pH adjustment step, the pH can be adjusted with, for example, an aqueous hydrochloric acid solution.

  In the pH adjustment step, the pH is preferably adjusted to 1 to 4, and more preferably adjusted to 1 to 3. By adjusting pH to 1-4, there exists an advantage that said compound can be extracted more reliably by an organic solvent for the same reason as mentioned above.

  In the organic solvent extraction step, the organic solvent extract is obtained by subjecting the pH-adjusted fraction to an extraction treatment with an organic solvent. That is, an organic solvent solution obtained by dissolving the above compound in an organic solvent is obtained as an extract.

In the organic solvent extraction step, a general extraction process can be adopted as the extraction process.
As the extraction process, for example, an extraction process using a separatory funnel or an extraction process using a Soxhlet extractor can be employed.

  In the organic solvent extraction step, a general organic solvent is used as the extraction solvent. In addition, the extraction solvent in an organic solvent extraction process normally contains 95 volume% or more of organic solvents.

  In the said organic solvent extraction process, extraction temperature is 20-30 degreeC normally. The extraction time is, for example, 30 minutes to 24 hours.

  Examples of the organic solvent include a polar organic solvent having a polar group in the molecule, or a nonpolar organic solvent having no polar group in the molecule.

  Examples of the polar organic solvent include those having an ester group, those having a hydroxy group, those having an ether group, those having a ketone group, and the like as polar groups.

Examples of the polar organic solvent having an ester group include carboxylic acid esters such as ethyl acetate and butyl acetate.
Examples of the polar organic solvent having a hydroxy group include monohydric alcohols such as methyl alcohol, ethyl alcohol, and butyl alcohol.
Examples of the polar organic solvent having an ether group include monoether compounds such as diethyl ether and tetrahydrofuran.
Examples of the polar organic solvent having a ketone group include acetone and methyl ethyl ketone.

  Examples of the nonpolar organic solvent include compounds having an aromatic hydrocarbon structure such as benzene, toluene and xylene.

  The organic solvent is preferably a polar organic solvent having 2 to 4 carbon atoms in that the above compound can be extracted more reliably, and is a carboxylic acid ester having 2 to 4 carbon atoms or a monohydric alcohol having 2 to 4 carbon atoms. Is more preferable, and ethyl acetate or ethyl alcohol is more preferable.

  In addition, the said organic-solvent extraction process can be performed using the water extract already extracted from the coffee bean with water. For example, commercially available instant coffee (a water extract of roasted coffee beans) can be employed as the water extract.

  At the said refinement | purification process, a refinement | purification process can be performed to the organic-solvent extract after the extraction process containing said compound by employ | adopting a general purification method.

  As the purification treatment, for example, a method performed in the above-described fractionation step can be employed. Further, a purification treatment by reverse osmosis, a purification treatment by adsorption, and the like can be employed.

  According to the method for producing a supercooling accelerator of the present embodiment, it is possible to obtain a supercooling accelerator that has both supercooling promoting ability and antioxidant performance without chemical synthesis. Accordingly, since such a supercooling accelerator can be obtained without using a synthesis catalyst or the like, it can be relatively safe for the human body.

  The supercooling accelerator produced as described above is blended in a composition containing at least water, and is suitably used, for example, in processed foods, cosmetics, and the like.

  Subsequently, an embodiment of the anticoagulant composition according to the present invention will be described in detail.

The anticoagulant composition of this embodiment contains at least the above-mentioned supercooling accelerator and water.
The anticoagulant composition usually further includes various compounding ingredients.

Since the anti-coagulant composition contains the supercooling accelerator, the formation of ice nuclei is suppressed even when the temperature is lower than the melting point of water. Therefore, according to the composition, even if the composition is placed in a sub-freezing environment for the purpose of storage or the like, it can be suppressed that the blended components blended in the composition are destroyed due to freezing.
Furthermore, since the said anticoagulant composition contains said supercooling promoter, it has sufficient antioxidant performance.
That is, the anticoagulant composition has a sufficient performance for causing a supercooling phenomenon and a sufficient antioxidant performance.

  Examples of the anticoagulant composition include liquid beverages, frozen foods, cosmetics, blood for testing, cell suspensions, organ preservation solutions, paints, and the like.

  The anticoagulant composition usually contains 0.01% by mass to 90% by mass of the above supercooling accelerator in terms of the above compound.

  The anticoagulant composition is suitably used in technical fields such as the food field, cosmetic field, medical field, and paint field.

  Furthermore, an embodiment of the method for producing an anticoagulant composition according to the present invention will be described in detail.

The method for producing an anticoagulant composition according to this embodiment includes a step of mixing the above-described supercooling accelerator and a liquid containing water (hereinafter also referred to as a mixing step).
According to the method for producing an anticoagulant composition of the present embodiment, it is possible to obtain a composition in which the formation of ice nuclei is suppressed even under a freezing temperature environment. Moreover, the composition which has antioxidant performance can be obtained.

  The mixing step is performed by a general method. By performing the mixing step, for example, an anticoagulant composition as described above can be obtained.

  The manufacturing method of the anticoagulant composition of this embodiment is used suitably in the technical field mentioned above, for example.

  The supercooling accelerator, the method for producing the supercooling accelerator, the anticoagulant composition, and the method for producing the anticoagulant composition of the present embodiment are as illustrated above, but the present invention is as illustrated above. The form is not limited. Further, in the present invention, various effects adopted in the general supercooling accelerator, the method for producing the supercooling accelerator, the anticoagulant composition, and the method for producing the anticoagulant composition are obtained according to the effects of the present invention. Can be employed within a range that does not impair the process.

  EXAMPLES Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

Example 1
A supercooling accelerator was produced as follows.
"Roasting process"
The seeds of plants (Arabica species) belonging to the genus Coffea were roasted at 180 ° C.
"Water extraction process"
A coffee koji was prepared after the roasted coffee beans were extracted with water at 85 ° C.
100 g (dry matter) of this coffee cake is put into 500 mL of deionized water and extracted in an autoclave at 90 ° C. for 1 hour, and the residue after extraction is removed by centrifugation (8000 g, 20 minutes). 5.7 g of water extract (water extract) was obtained.
"Fractionation process"
Fractionation was performed by filtering the obtained liquid water extract with an ultrafiltration membrane.
In the fractionation treatment with an ultrafiltration membrane, an ultrafiltration device equipped with an ultrafiltration membrane having a molecular weight cut off of 10,000 (manufactured by Nihon Millipore, ultrafiltration disk, ultracell, PL, regenerated cellulose, 10000NMWL) Amicon Stirring Cell Model 18400 manufactured by Nippon Millipore was used.
A fraction solution having a molecular weight of 10,000 or less was obtained.
"PH adjustment process"
The pH was adjusted to 2.0 by adding an aqueous hydrochloric acid solution to the fraction solution after fractionation.
"Organic solvent extraction process"
The organic solvent extraction process was performed by putting the fraction liquid after adjusting pH and ethyl acetate in a separating funnel and shaking the separating funnel.
Then, the ethyl acetate layer was taken out and the ethyl acetate was volatilized to obtain a solid crude extract. Such a crude extract was obtained as a supercooling accelerator.
In addition, in the “evaluation of supercooling promotion ability (anti-ice nucleus activity ability)” described later, a residue after removing water by volatilizing and removing it from the aqueous layer in the organic solvent extraction step was used as a comparative sample.

(Example 2)
Using the supercooling accelerator produced in Example 1, a purification process was further performed.
"Purification process"
That is, the purification process was performed by anion exchange chromatography and gel filtration chromatography.
Anion exchange chromatography 40 mg of the above crude extract was diluted with 50 ml of Tris-HCl buffer (10 mM, pH 8.0) to obtain a purification solution.
50 ml of this solution was purified by anion exchange chromatography under the following conditions.
Column: DEAE column equilibrated with buffer
(DEAE-TOYOPEARL650M, 16 mm × 20 cm,
Manufactured by Tosoh Corporation)
Flow rate: 2.0 ml / min Concentration gradient: 20 mM to 1 M Ammonium acetate Total elution volume: 400 ml
Then, a fraction eluted at an ammonium acetate concentration of around 0.4 M (elution amount of around 180 ml) was collected.
Gel filtration chromatography The fraction obtained by anion exchange chromatography was further purified by subjecting it to gel filtration chromatography under the following conditions.
Carrier: Sephadex LH-20 (manufactured by GE Healthcare)
Eluent: 50% by volume of methanol Column: φ15mm x 1000mm
Flow rate: 1.0 ml / min Apparatus: Flash chromatograph SYS16020 (manufactured by Tokyo Science Instruments Co., Ltd.)
Liquid volume: 300ml
Then, 3 ml of fraction eluted at around 100 ml of eluate was collected.

The liquid supercooling accelerator of Example 1 was produced as described above.
An anion exchange chromatography chart is shown in FIG. 1, and a gel filtration chromatography chart is shown in FIG.

<Analysis of compounds contained in supercooling accelerator>
About the compound contained in the supercooling promoter manufactured in Example 2, the molecular structure was analyzed by the conventional method.
That is, the compound was analyzed by LC-MS, MS-MS, ¹ H-NMR.
Conditions in each analysis are as follows.
LC-MS: Analytical instrument-"API3000" manufactured by Applied Biosystems
HPLC instrument-Agilent "1100series"
Mobile phase-acetonitrile / water, flow rate 1.0 ml / min
Column size -4.6mm x 150mm
MS-MS: Atmospheric pressure chemical ionization-mass spectrometer (APCI-MSMS)
(Analytical instrument-"API3000" manufactured by Applied Biosystems)
(Measurement condition-mobile phase 1% formic acid ultrapure aqueous solution and 1% formic acid acetonitrile solution were mixed at 1: 1 and analyzed with Positive (M + 1) at a flow rate of 1 ml / ml)
¹ H-NMR: Analytical instrument “JEOL EX-400” manufactured by JEOL Ltd. (399.65 MHz, D ₂ O)
FIG. 3 shows the results of LC-MS, FIG. 4 shows the results of MS-MS, and FIG. 5 shows the results of ¹ H-NMR.
From these results, it was found that the supercooling accelerators of Example 1 and Example 2 contained at least a compound having an aromatic hydrocarbon structure and a carboxy group in the molecule. Further, it was found that such compounds in Example 1 and Example 2 had a composition formula of C ₁₅ H ₂₆ N ₂ O ₇ and a molecular weight of 346.

(Example 3)
A supercooling accelerator was produced as follows.
Specifically, first, 4 g of instant coffee (Blendy AGF) dissolved in 200 ml of ultrapure water (20 mg / ml) was prepared. The pH was then adjusted to 2.0 with 1M HCl. Then, the organic solvent extraction process was performed with 2 volumes of ethyl acetate using a separatory funnel. The ethyl acetate layer after fractionation was collected and concentrated to dryness using an evaporator to produce a solid supercooling accelerator. The recovery rate of the supercooling accelerator was 1.5 g (37.5%).
The molecular structure of the compound contained in the supercooling accelerator of Example 3 was analyzed in the same manner as described above. As a result, it was found that the compound contained in the supercooling accelerator of Example 3 had at least an aromatic hydrocarbon structure and a carboxy group in the molecule.

<Evaluation of supercooling promotion ability (anti-ice nucleus activity ability)>
With respect to the supercooling accelerator of Example 1 and Example 3, the supercooling promoting ability was evaluated by the following method.

First, 150 mM phosphoric acid so that the solid content concentration of the crude extract (residue) is 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 5.0 mg / mL, 10.0 mg / mL. The solution was diluted with a buffer solution (pH 7.0), and the diluted solution was filtered with a filter (Cellulose Acetate 0.2 μm manufactured by ADVANTEC). 100 μl of each aqueous solution after filtration was prepared.
Meanwhile, 900 μl of an aqueous solution in which silver iodide was dissolved in 150 mM phosphate buffer (pH 7.0 KPB) was prepared so that the silver iodide concentration was 1 mg / ml.
Each sample for evaluation was prepared by mixing the above two aqueous solutions.
In addition, what was prepared similarly using the residue from the aqueous layer obtained in Example 1 as a sample for comparative evaluation was prepared. Moreover, what mixed 100 microliters of 150 mM phosphate buffers and said silver iodide solution was used for the blank sample.
Supercooling promotion ability (anti-ice nucleus activity ability) was measured by Vali's droplet freezing method. Specifically, in Vali's droplet freezing method, an aluminum film is placed on a copper plate of a cold plate cooling device (COOLACE CCA-1000 EYELA), and 10 μL of each sample for evaluation and blank sample are placed on the surface. It is dripped at a spot, the temperature is lowered at a rate of 1.0 ° C. per minute, and T ₅₀ is a temperature at which 50% of 30 droplets are frozen.
The temperature during the droplet freezing of the sample and Samplet _50, the temperature during the droplet freezing of blank sample and BlankT _50. Then, the anti-ice nucleation activity values [Delta] T ₅₀ (° C.) was determined by the following equation.
ΔT ₅₀ (° C.) = BlankT ₅₀ -SampleT ₅₀

The results of evaluating the supercooling accelerator of Example 1 are shown in FIG. The result of evaluating the supercooling accelerator of Example 3 is shown in FIG. In addition, the value of a result is an average value of 3 times of test results.
As understood from FIGS. 6 and 7, the supercooling accelerator of Example 1 was excellent in supercooling promoting ability (anti-ice nucleus activity ability).

Regarding the supercooling accelerator of Example 2, the antioxidant performance was evaluated by the following method.

<Evaluation of antioxidant performance>
The antioxidant performance was evaluated by a generally known diphenylpicrylhydrazyl (DPPH) radical scavenging activity test.
Specifically, in a 1.5 ml sampling tube, ultrapure water (250 μl), ethanol (300 μl), 200 mM MES aqueous solution (300 μl), and the supercooling accelerator of Example 2 were prepared to have a predetermined concentration in terms of solid content. The aqueous solution (50 μl) and 400 μM DPPH ethanol solution (300 μl) were added sequentially.
Then, the contents were stirred with a vortex mixer. Then, it left still for 20 minutes in the dark place at room temperature. Thereafter, the absorbance of the sample for evaluation was measured at a wavelength of 520 nm using an absorptiometer. As a blank sample, ultrapure water (50 μl) was used.
DPPH radical scavenging activity (%) was calculated by the following formula.
DPPH radical scavenging activity (%) = {1− (A ₅₂₀ / A ′ ₅₂₀ )} × 100
[A ₅₂₀ = absorbance of sample for evaluation A ' ₅₂₀ = absorbance of blank sample]

The evaluation results of the antioxidant performance are shown in FIG. As can be seen from FIG. 8, the supercooling accelerator of Example 2 was excellent in antioxidant performance.
In addition, about the supercooling promoter of Example 2, when antioxidant activity was evaluated by Trolox equivalent, it was 6.72 micromol TE / g (solid content conversion).

  The supercooling accelerator of the present invention is suitably used for the purpose of suppressing freezing of a composition containing the supercooling accelerator and water, for example, in the processed food field, cosmetic field, and medical field. Similarly, the anticoagulant composition of the present invention is suitably used, for example, in the processed food field, the cosmetic field, and the medical field for the purpose of suppressing the freezing of the liquid composition below freezing.

Claims (3)

A supercooling accelerator comprising a compound represented by a composition formula of C ₁₅ H ₂₆ N ₂ O ₇ extracted from coffee beans, wherein the compound has at least an aromatic hydrocarbon structure and a carboxy group in the molecule.   The supercooling accelerator according to claim 1, wherein the coffee beans are roasted coffee beans. A method of manufacturing a supercooling promoting agent comprising at least an aromatic and a hydrocarbon structure and a carboxy group C ₁₅ H ₂₆ N ₂ O ₇ of Compositional Formula compound in the molecule,
A water extraction step of obtaining a water extract by subjecting the coffee beans to extraction with water;
A fractionation step of obtaining a fraction solution containing the compound by subjecting the water extract extracted by the water extraction step to a fractionation treatment according to the molecular weight;
A pH adjusting step for adjusting the pH of the fractionated liquid containing the compound to the acidic side;
An organic solvent extraction step of performing extraction with an organic solvent on the pH-adjusted fraction solution, and a method for producing a supercooling accelerator.