JP2004309454A - Indicator substance for determining body odor, handy body odor determining kit, method for determining body odor, and method for determining effectiveness of deodorant - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an indicator substance capable of objectively and quantitatively determining the presence or absence and strength of especially apocrine odor, of which many people are conscious, among various kinds of body odors, to provide a method for determining the degree of body odor or the effectiveness of deodorant by using the indicator substance; to provide a body odor determination kit for accurately, quickly, and simply determining the type of body odor and the degree of odor, and to provide a method for determining the body odor by using the body odor determination kit. <P>SOLUTION: The indicator substance for determining body odor contains a β-hydroxy acid compound (1) expressed by formula (1), and at least one selected from a group made of the indicator compound. In the expression, R<SP>1</SP>, R<SP>2</SP>indicate alkyl having 1-4 carbons, and hydrogen atom or alkyl having 1-4 carbons, and the number of carbons in the formula (1) is equal to or less than 10. Additionally, the body odor determining kit contains a color identification reagent reacting with a fatty acid, where the number of carbons of those other than β-hydroxycarboxylic acid and/or β-hydroxycarboxylic acid originating in human sweat is 12 or less. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

The present invention relates to an indicator substance that can be used for determining body odor, and a method for determining the degree of body odor or the effectiveness of a deodorant using the indicator substance.
In the present invention, human sweat-derived β-hydroxycarboxylic acid or the like is reacted with a coloring agent, and the type of body odor (acid odor, apocrine odor, mixed odor) and the intensity of the body odor are easily determined based on the developed color. The present invention also relates to a determination kit and a determination method capable of performing the determination.

  In recent years, people who are concerned about body odor are increasing with an increasing tendency toward cleanliness. Body odor is a general term for odors generated from various parts of the whole body, and the main sites of occurrence include the head, oral cavity, axilla, genital area, soles of the feet, and the like. Above all, since the armpit odor is easily perceived by the person or the person who is nearby, various factors such as the presence or absence of the armpit odor, the strength, and the qualitative difference often control the degree of the body odor of the whole body. Furthermore, for the person or the person who is nearby, there are many cases where the axillary odor itself is more noticeable than the body odor.

  In the human axilla, not only eccrine sweat glands distributed throughout the whole body but also special sweat glands called apocrine sweat glands are present. The axillary region is a site where sweat is difficult to evaporate, and because it is also a site where bacteria easily grow, sweat (eccrine sweat and apocrine sweat) secreted from these two sweat glands, sebum, dirt, etc. on the skin It is easily metabolized by bacteria and produces odor.

  The odor derived from eccrine sweat is commonly called acid odor or sweat odor, and is generated not only from the armpit but also from the skin surface of the whole body. On the other hand, the odor derived from the apocrine sweat glands is generated from the so-called axillary part of a person who is called aside, and is distinguished from apocrine odor or simply called "armpit odor".

  Axillary odor has individual differences, and is roughly classified into acid odor, apocrine odor, and a mixed odor of both. The acid odor is generally an odor caused by a lower carboxylic acid having 2 to 5 carbon atoms, and the apocrine odor is an odor peculiar to the axilla (so-called “sidearm”). Apocrine gland secretions (apocrine sweat) are essentially odorless by nature, but are known to be metabolized by bacteria on the skin after secretion to produce an apocrine odor.

  People with aside tend to have a large number of apocrine sweat glands in the axilla, and the sweat odor generated by eccrine sweat and the apocrine odor mix to form a strong and unique odor. Therefore, if you are worried about your own armpit odor, use a deodorant with antiperspirant or bactericidal action, or if the apocrine odor is extremely strong, surgically remove the apocrine sweat glands in the axilla. Attempts are made to reduce the smell.

  For those who are worried about body odor or armpit odor, and even at a level that you do not notice, you are in a occupation where the armpits are close to the face of another person like a hairdresser or dentist For people, people who have pointed out body odor at home, school, company, etc., how much apocrine odor originally exists in their own axillary area, use of deodorants and removal surgery for apocrine sweat glands It is an important concern whether efforts to reduce body odor, etc. have been successful in the current situation.

Conventionally, methods for determining axillary odor include (1) a sensory evaluation test in which a third party sniffs the odor of sweat in the axillary region with the nose, (2) there is a person with an armpit in the family, and the earwax is wet. Empirical determination methods by investigating the fact that the relevance of the underwear to the axillary part is colored by sweat, and (3) a method of estimating from the number and size of apocrine sweat glands.

  The method (1) first requires a specialized panelist such as a skilled dermatologist and cannot be easily implemented. In addition, this method wipes off the smell of the test subject's armpit with absorbent cotton, etc., and sensuously determines the degree of apocrine odor.However, there is much room for the subjective evaluation of the evaluator, and quantitative evaluation is not possible. difficult. In the case of continuous evaluation, the objectivity may be reduced due to olfactory fatigue.

The investigation of the related facts in (2) is an indirect judgment based on a judgment based on genetic predisposition and a correlation with the degree of wetness of earwax. Further, the judgment based on the coloring of the underwear focuses on the pigment contained in the sweat of the apocrine gland, but does not directly evaluate the apocrine odor in the axilla.
Therefore, within the range of the evaluation items, there is a possibility that an erroneous determination may be made, and there is a possibility that an underarm odor that does not become apparent may be missed. In addition, such a survey can be used as a guide to determine the presence or absence of axillary odor, but because it is non-quantitative and inaccurate, the situation of improvement of axillary odor due to apocrine gland removal surgery and regeneration of apocrine gland after surgery It is not practical to judge the recurrence and degree of apocrine odor due to odor, the degree of the masking effect by the deodorant, and the like.

  The method (3) is a method in which a doctor surgically incises the axilla to observe the number and size of the apocrine sweat glands and estimate the degree of body odor from the appearance. It is not easy to implement because of great pain. In general, surgery costs are high.

  Therefore, among the body odors, there are many people who particularly care about the apocrine odor, and it is an indicator substance that enables objective and quantitative determination of the presence and strength of the apocrine odor, and the degree of the body odor or the effectiveness of the deodorant agent using it. It is desired to provide a method for judging sex, and a judgment kit and a judgment method capable of diagnosing body odor, particularly apocrine odor, reliably, quickly and easily.

  Non-Patent Document 1 discloses that underarm sweat includes, in addition to lower carboxylic acid contained in normal sweat secreted from the soles of the soles and the skin of the chest, trans as a characteristic causative component of apocrine odor under the armpits There is a description that -3-methyl-2-hexenoic acid, 7-octenoic acid and the like are contained.

However, in the axilla of human beings, not only the components causing the apocrine odor, but also lower carboxylic acids having 5 or less carbon atoms and higher carboxylic acids derived from sebum, which cause the acid odor, are mixed. It was difficult to selectively separate only the aforementioned unsaturated carboxylic acid, which is a characteristic component of the apocrine odor, from the components.
In addition, Patent Literature 1 describes that a specific β-hydroxycarboxylic acid or a salt thereof is used as a material for an animal flavor, but the relationship between this compound and axillary odor is not known.

JP-A-10-25265 "Molecular Recognition of Taste and Smell", Review of Chemistry No.40, 1999, p.205-211

  An object of the present invention is to provide an indicator substance capable of objectively and quantitatively determining the presence and intensity of apocrine odor, which is often noticeable among body odors, and the degree of body odor or It is an object of the present invention to provide a method for determining the effectiveness of a deodorant agent, a body odor determination kit capable of reliably and quickly and easily determining the type and degree of body odor, and a method for determining body odor using the kit. .

  The present inventors have conducted intensive studies on the causative component of apocrine odor contained in axillary sweat and found that it is very similar to apocrine odor, specifically present in sweat of a person with apocrine odor, and quantified. Has a concentration that can be determined by simple chemical manipulations and found that a component that can be isolated from sweat is present in the sweat, which objectively determines the degree of apocrine odor in the axilla. Could be used as an important indicator.

That is, since the present inventors detected and identified 3-hydroxy-3-methylhexanoic acid from underarm sweat, it is a major causative substance of apocrine odor in the axillary region, and as an objective index for determining apocrine odor. Can be used.
Also, a β-hydroxy acid compound similar to 3-hydroxy-3-methylhexanoic acid is used as an objective index for determining apocrine odor, like 3-hydroxy-3-methylhexanoic acid. be able to.

  Furthermore, the present inventors separated β-hydroxycarboxylic acid containing 3-hydroxy-3-methylhexanoic acid, which was newly found as a major causative substance of apocrine odor, from axillary sweat, and then developed a coloring reagent. Focusing on the fact that the degree of apocrine odor can be easily determined by the developed color, the degree of apocrine odor itself, or the degree of total body odor centered on apocrine odor can be reliably and easily determined. A kit capable of determination and a method for determining body odor using the kit have been developed.

  The present invention is an indicator substance for body odor determination containing at least one selected from the group consisting of β-hydroxy acid compounds represented by the following formula (1).

（式中、Ｒ¹は炭素数１乃至４のアルキルであり、Ｒ²は水素原子又は炭素数１乃至４のアルキルであり、式（１）の総炭素数が１０以下である。）

(In the formula, R ¹ is an alkyl having 1 to 4 carbons, R ² is a hydrogen atom or an alkyl having 1 to 4 carbons, and the total number of carbons in the formula (1) is 10 or less.)

  Further, the present invention provides a body odor containing a β-hydroxy acid derivative obtained by introducing an atom or an atomic group into a β-hydroxyl group and / or a carboxyl group of a β-hydroxy acid compound represented by the above formula (1). It is also an indicator substance for judgment. In order to improve analytical sensitivity in instrumental analysis, β-hydroxy acid can be derivatized, or a labeling substance can be introduced into β-hydroxy acid. The above-mentioned indicator substance is suitably used for determining body odor and determining the effectiveness of a deodorant agent.

  The present invention is also a human body odor determination kit including a color reagent which reacts with human sweat-derived β-hydroxycarboxylic acid and / or a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid. The degree of body odor derived from human sweat is determined by using a β-hydroxycarboxylic acid which is a causative substance of apocrine odor, and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid which is a causative substance of acid odor. The odor type and the intensity of the odor can be reliably, quickly and easily determined by the developed color.

The first method of determining body odor using the kit of the present invention includes a first step of extracting a mixture of β-hydroxycarboxylic acid and fatty acids having 12 or less carbon atoms other than the β-hydroxycarboxylic acid contained in human sweat. ,
A second step of adding the reagent of the kit to the mixture to form a color,
A third step of determining the type and / or intensity of the body odor based on the color developed in the second step.

In addition, the second method for determining body odor using the kit of the present invention is a method for extracting a mixture of β-hydroxycarboxylic acid and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid contained in human sweat. Steps and
A second step of separating β-hydroxycarboxylic acid from the mixture;
A third step of reacting the reagent with the β-hydroxycarboxylic acid separated in the second step to form a color,
A fourth step of determining the type and / or intensity of the body odor based on the color developed in the third step.

Further, the third method for judging body odor using the kit of the present invention is a method for extracting a mixture of β-hydroxycarboxylic acid contained in human sweat and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid. Steps and
A second step of separating the mixture into a β-hydroxycarboxylic acid and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid,
A third step of reacting the reagent of the kit with the β-hydroxycarboxylic acid separated in the second step to form a color,
A fourth step of reacting the reagent of the kit with a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid separated in the second step to form a color,
A fifth step of determining the type and / or intensity of the body odor based on the colors developed in the third and fourth steps.

  Due to the indicator substance containing at least one selected from the group consisting of the β-hydroxy acid compound (1) represented by the formula (1) according to the present invention and its labeled compound, it is a human body odor or a part of body odor. The degree of axillary odor can be objectively and quantitatively determined based on the presence or absence and strength of apocrine odor in the axillary region.

  In addition, the indicator substance for body odor judgment of the present invention expresses the subjectivity from the judgment result by expressing the measured value by the amount of 3-hydroxy-3-methylhexanoic acid, particularly by chemical analysis or instrumental analysis. It is possible to eliminate.

  Also, despite the large amount of 3-hydroxy-3-methylhexanoic acid produced in the axilla, the latent state of axillary odor, as in the case where it has changed to a weak or weak derivative such as salt, etc. Also, in the present invention, 3-hydroxy-3-methylhexanoic acid can be detected and directly evaluated by directly measuring it.

  Further, in the present invention, the effectiveness of a deodorant agent targeting an apocrine odor can be objectively and quantitatively determined using an indicator substance containing the β-hydroxy acid compound (1) or a derivative thereof.

The kit for determining body odor according to the present invention comprises a β-hydroxycarboxylic acid specifically present in sweat of a person having an apocrine odor and / or a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid which is a causative substance of the acid odor. Or, utilizing the color reaction of its derivative or its decomposed product, the degree of axillary odor, which is a part of human body odor or body odor, is centered on the presence and intensity of apocrine odor in the axilla, and the presence or absence and intensity of acid odor In consideration of the above, the determination can be made reliably, quickly and easily.
In addition, the body odor determination kit and the determination method of the present invention can eliminate the subjectivity from the determination result by expressing the hue expressed by the color reaction by numerical values using an analyzer such as a color difference meter. is there.

  Furthermore, the body odor determination kit and the determination method of the present invention can be used in hospitals, public health centers, research laboratories, and the like, when diagnosing the degree of apocrine odor for a person who is interested in own axillary odor, the effect of removing apocrine sweat glands and the like. Can be used as an accurate, quick and simple body odor determination kit and determination method when evaluating the effectiveness of a deodorant agent sample targeting an apocrine odor.

  The indicator substance for body odor determination according to the present invention contains at least one selected from the group consisting of β-hydroxy acid compounds represented by the following formula (1) (hereinafter referred to as β-hydroxy acid compounds (1)). Alternatively, the β-hydroxy acid compound (1) may be used alone.

（式中、Ｒ¹は炭素数１乃至４のアルキルであり、Ｒ²は水素原子又は炭素数１乃至４のアルキルであり、式（１）の総炭素数が１０以下である。）

(In the formula, R ¹ is an alkyl having 1 to 4 carbons, R ² is a hydrogen atom or an alkyl having 1 to 4 carbons, and the total number of carbons in the formula (1) is 10 or less.)

In the above chemical formula (1), R ¹ is an alkyl having 1 to 4 carbons, and the alkyl having 1 to 4 carbons may be any of linear or branched alkyl, for example, methyl, ethyl, n-propyl , I-propyl, n-butyl, i-butyl and t-butyl. Since the β-hydroxy acid compound (1) is considered to be easier to use as an index substance as the property of 3-hydroxy-3-methylhexanoic acid, which is the object of detection, is closer, the chemical structure of the compound is represented by 3-hydroxy-3- In order to approach methylhexanoic acid, R ¹ preferably has 3 or 4 carbon atoms, particularly 3 carbon atoms, and is preferably straight-chain alkyl.

In the above chemical formula (1), R ² is a hydrogen atom or an alkyl having 1 to 4 carbon atoms, and the alkyl may be any of linear or branched alkyl. The R ^2, for example, methyl, ethyl, n- propyl, i- propyl, n- butyl, i- butyl, t- butyl. In order to bring the chemical structure of the β-hydroxy acid compound (1) closer to that of 3-hydroxy-3-methylhexanoic acid, the number of carbon atoms of R ² is preferably 1 or 2, particularly preferably 1.

  The β-hydroxy acid compound (1) is 3-hydroxy-3-methylhexanoic acid having an odor very similar to the apocrine odor, and a group of compounds very similar in chemical structure thereto. 3-hydroxy-3-methylhexanoic acid is represented by the following formula (2).

  3-Hydroxy-3-methylhexanoic acid is a known compound, and it is described in Patent Document 1 that it can be used as an animal-based synthetic fragrance material, but it is a major cause component of apocrine odor. Never before reported, such a fact was discovered by the present inventors.

  Since the above-mentioned 3-hydroxy-3-methylhexanoic acid discovered by the present inventors has the following features, the amount and the state of 3-hydroxy-3-methylhexanoic acid in the axilla are limited. It is thought that the degree of apocrine odor in the axilla and individual differences are formed.

(Characteristics of 3-hydroxy-3-methylhexanoic acid)
(1) A person who does not detect 3-hydroxy-3-methylhexanoic acid from axillary sweat has no apocrine odor, and 3-hydroxy-3-methylhexanoic acid is detected from axillary sweat One has an apocrine odor. That is, 3-hydroxy-3-methylhexanoic acid is specifically present in a person with an apocrine odor (FIGS. 1 and 2).
(2) The greater the amount of 3-hydroxy-3-methylhexanoic acid contained in sweat in the axilla, the stronger the apocrine odor (FIG. 3).

(3) Since the β-hydroxy acid compound (1) has not only a carboxyl group but also a β-hydroxyl group, it is easy to perform chemical modification for highly sensitive detection in instrumental analysis. In addition, chemical properties often differ from ordinary carboxylic acids, and by utilizing differences in solubility, etc., hydroxy acids and / or derivatives thereof and other lower carboxylic acids having low contribution to apocrine odor are used. Separation from acid is facilitated.
(4) Since 3-hydroxy-3-methylhexanoic acid has a hydroxyl group, it has a high molecular polarity and can be isolated by a method such as adsorption column chromatography. Quantitative evaluation can be performed using a color reaction.

  Therefore, the amount of 3-hydroxy-3-methylhexanoic acid and / or its derivative in the axilla and its state (eg, 3-hydroxy-3-methylhexanoate or ester) can be determined chemically or physically. Measurement by an appropriate method such as the above means that the apocrine odor in the axilla is objectively and quantitatively measured using 3-hydroxy-3-methylhexanoic acid as an index.

Further, the β-hydroxy acid compound (1) other than 3-hydroxy-3-methylhexanoic acid has a chemical structure and chemical and functional properties (particularly, 3-hydroxy-3-methylhexanoic acid) Since properties such as odor are similar, it can be used as an objective index for determining apocrine odor, similarly to 3-hydroxy-3-methylhexanoic acid.
Among the β-hydroxy acid compounds (1), 3-hydroxy-3-methylhexanoic acid is particularly suitable as an indicator substance because it is the main causative component of apocrine odor present in axillary sweat.

  The β-hydroxy acid compound (1) may be used after being chemically modified as long as the detection function as an indicator compound is not lost. For example, a derivative in which one or both of a carboxyl group and a β-position hydroxyl group are converted to a salt, ester, amide, ether, or the like can be used.

  As derivatization reagents, UV reagents such as O- (p-nitrobenzyl) -N, N '-(diisopropylisourea) (PNBDI) and p-bromophenacyl bromide (PBPB), and 4-bromomethyl-7- Fluorescent reagents such as methoxycoumarin (Br-MmC), silylating agents such as N-trimethylsilylimidazole (TMSI) and N, O-bis (trimethylsilyl) acetamide (BSA), and acyls such as trifluoroacetic anhydride and trifluoroacetylimidazole An agent or the like can be used.

  When a chromophore in the visible region is used as the labeled compound of the β-hydroxy acid compound (1), a concentration-color standard sample of the labeled compound is prepared, and sweat collected from a human is colored with the same reagent. It is also possible to visually judge the degree of apocrine odor as compared with.

  The β-hydroxy acid compound (1) can be synthesized, and is suitable as an indicator substance in that the apocrine odor can be objectively evaluated and determined at any time and place by stably supplying a synthetic product of constant quality. I have. The β-hydroxy acid compound (1) can be prepared, for example, according to the following reaction formula (3) by a Reformatsky reaction [Reformatsky Reaction; Ber. 20, 1210 (1887), J. Russ. Phys. Chem. Soc., 22, 44 ( 1890)] to synthesize an ester having a hydroxyl group at the β-position, and hydrolyze the ester to synthesize the ester.

（式中、Ｒ¹及びＲ²は前記と同じであり、Ｘはハロゲン原子である。）

(In the formula, R ¹ and R ² are the same as above, and X is a halogen atom.)

The synthesized β-hydroxy acid compound (1) can be appropriately converted to a salt, ester or other derivative by a known method, and may be used as a labeled compound.
In the present invention, in order to objectively and quantitatively determine the degree of axillary odor, which is a part of human body odor or body odor, from the presence or absence and strength of apocrine odor in the axillary region, synthesized β-hydroxy acid Using compound (1) or a derivative thereof, the amount of 3-hydroxy-3-methylhexanoic acid contained in the sweat in the axilla is quantified, and the state of the presence is observed.

  The method of using the β-hydroxy acid compound (1) or a derivative thereof as an indicator substance for body odor determination is not particularly limited, and may be used in accordance with various known evaluation methods. For example, when the content of 3-hydroxy-3-methylhexanoic acid contained in sweat in the axillary region is measured by GC-MS, the indicator substance of the present invention, preferably 3-hydroxy-3- as an active ingredient, is used. A calibration curve is prepared using methylhexanoic acid or a derivative thereof as a standard substance (standard). Using this calibration curve, the peak of 3-hydroxy-3-methylhexanoic acid contained in the collected sweat may be identified and its amount may be measured.

  In the case of performing a sensory evaluation, the indicator substance of the present invention, preferably 3-hydroxy-3-methylhexanoic acid or a derivative thereof is used as an active ingredient, diluted in several steps, and an odor standard sample of each concentration is prepared. Prepare. Then, the odor of the test sample prepared from the sweat collected from the axilla is compared with the standard sample, and the amount of 3-hydroxy-3-methylhexanoic acid contained in the sweat may be determined by sensory evaluation.

  As a measuring method for determining body odor, not only the indicator substance itself according to the present invention can be used alone, but also a derivatized version of the indicator substance can be used. Further, after the test paper is impregnated with the indicator substance, it may be used directly for determination.

  The visual sensory evaluation includes, for example, an evaluation of visually judging the color of the aqueous solution, and an evaluation using a test paper impregnated with a labeling compound.

  In this manner, the degree of axocrine odor in the axillary region is quantitatively determined using the odor or a parameter other than the odor, thereby determining the degree of the body odor or the part of the body odor.

  In addition, when a large amount of 3-hydroxy-3-methylhexanoic acid is produced in the axilla, but it has changed to an odorless or weak derivative such as a salt, a latent state of axillary odor exists. However, there are cases where such a state cannot be accurately evaluated even by conducting a sensory evaluation or investigating a fact that is related to the armpit odor. On the other hand, in the present invention, by measuring 3-hydroxy-3-methylhexanoic acid or a derivative thereof that can be analyzed by the necessary chemical treatment, whether or not the subject has a constitution that may cause axillary odor is determined. That is, potential evaluation can be performed.

  The indicator substance of the present invention can be used for any of chemical analysis, instrumental analysis, sensory evaluation, and the like as described above, and enables highly objective quantitative determination.In particular, measurement is performed by chemical analysis or instrumental analysis. By expressing the value by the amount of 3-hydroxy-3-methylhexanoic acid, subjectivity can be excluded from the determination result.

  Further, in the present invention, the effectiveness of a deodorant agent targeting an apocrine odor can be objectively and quantitatively determined using an indicator substance containing the β-hydroxy acid compound (1) or a derivative thereof.

  As a method of determining the effectiveness of the deodorant, the indicator substance may be used alone, and other components such as a solvent for dissolution or dilution, a stabilizer, an antiperspirant, a bactericide, and an antibacterial agent Further, additives such as a surfactant, an antioxidant, a fragrance, and a plant extract may be blended to prepare a composition suitable for practical use such as storage and use in a judgment test.

  A deodorant agent targeting an apocrine odor can be any type such as a type that sterilizes bacteria on the skin and prevents the decomposition of sweat, a type that decomposes or changes odor components into odorless derivatives, or a type that masks odors A type of action mechanism may be used. The method of using the β-hydroxy acid compound (1) or a derivative thereof as an indicator for determining the effectiveness of a deodorant agent is not particularly limited, and may be used according to the mechanism of action and the evaluation method of the deodorant agent.

  For example, a predetermined amount of a deodorant agent sample is added to an indicator substance containing a β-hydroxy acid compound (1) or a derivative thereof, preferably 3-hydroxy-3-methylhexanoic acid or a derivative thereof at a predetermined concentration as an active ingredient. Then, by quantifying the change state of the indicator substance by an appropriate method, the effectiveness of the deodorant sample can be objectively and quantitatively determined.

  As a method of quantifying the change state of the indicator substance, when the deodorant sample is of a type that degrades 3-hydroxy-3-methylhexanoic acid or induces it to another compound to reduce the odor, A calibration curve may be prepared in advance, and instrumental analysis may be performed using this calibration curve, or a changed or unchanged indicator substance may be quantified by chemical analysis such as titration or extraction. If the deodorant sample is of the type that masks the apocrine odor, dilute the indicator in several steps to prepare odor standard samples at each concentration, and compare the odor of the indicator with the deodorant sample to the standard sample. Then, the masking effect may be determined by sensory evaluation.

  Further, as a labeling compound of the β-hydroxy acid compound (1), for example, a fluorescent labeling compound is used, and a predetermined amount of a deodorant agent sample is added to a labeling substance containing the labeling compound at a predetermined concentration, and the change state of the labeling substance is determined. The determination may be performed by performing instrumental analysis using a calibration curve of the same indicator substance. In addition, after adding a predetermined amount of a deodorant agent sample to the indicator substance containing the labeling compound at a predetermined concentration, and then quantifying the changed or unaltered form of the indicator substance by chemical analysis such as titration or extraction, the labeled part is used. It may be detected by utilizing.

  Furthermore, it is also possible to actually apply the deodorant sample to the human axilla, and to evaluate and compare the axillary sweat collected before and after the application using the indicator substance according to the present invention.

  In this manner, the apocrine odor of the indicator substance of the present invention caused by the action of the deodorant agent is quantitatively determined using an odor or a parameter other than the odor, so that the effectiveness of the deodorant agent is objectively and quantitatively determined. Can be evaluated.

The kit for determining body odor according to the present invention comprises at least a β-hydroxycarboxylic acid derived from human sweat and / or a coloring reagent that reacts with other body odor-causing components derived from sweat, and the β-hydroxycarboxylic acid and / or It is a product in which an accessory for assisting the determination based on the reaction and the coloration of the other body odor-causing component derived from sweat and the color reagent is included. There are essential ones such as an extraction unit used for preprocessing always performed in advance, and one that enhances convenience such as facilitating and simplifying a color reaction or determination.
The kit for determining body odor includes, for example, equipment and reagents for extracting and separating β-hydroxycarboxylic acid and / or fatty acids having 12 or less carbon atoms other than β-hydroxycarboxylic acid from axillary sweat, and Contains at least one reagent that develops a color by reacting with β-hydroxycarboxylic acid derived from perspiration and / or a fatty acid having 12 or less carbon atoms other than β-hydroxycarboxylic acid. Further, the kit may contain 3-hydroxy-3-methylhexanoic acid as an indicator component.
FIG. 4 shows an outline of a procedure for determining body odor derived from human sweat, particularly apocrine odor, by utilizing the color reaction of β-hydroxycarboxylic acid and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid. . 5 and 6 show typical equipment included in the kit for determining body odor and examples of its use.

  As a method of collecting sweat in the axilla, a method of directly collecting sweat that has flowed out due to heat sweating into a test tube, a T-shirt in which a cotton pad is sewn on a portion under the armpit, or a gauze is used. For example, there is a method of directly wiping the sweat under the armpit.

  The method for extracting the acidic substance contained in the sweat is not particularly limited as long as the acidic substance can be extracted, but an acid-base extraction method using an alkaline aqueous solution is generally used. Examples of the alkaline aqueous solution include an aqueous sodium hydrogen carbonate solution, an aqueous sodium carbonate solution, an aqueous sodium hydroxide solution, and an aqueous potassium hydroxide solution. Further, the acidic substance can be extracted using an ion exchange resin.

  As a method of determining the degree of apocrine odor, the acidic substance extracted by the above operation is separated into β-hydroxycarboxylic acid and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid, and then separated. A method of adding a color reagent to the β-hydroxycarboxylic acid and observing the developed color.

  The method for separating the β-hydroxycarboxylic acid and the fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid in the acidic substance is not particularly limited as long as it can be separated depending on the polarity of the molecule. Adsorption chromatography in which a suitable adsorbent such as silica gel powder is filled in a tube or a plastic tube, thin layer chromatography in which a suitable adsorbent such as silica gel is coated on a glass or plastic plate, or the like can be used.

  In the adsorption column chromatography method, when the solution of the acidic substance is allowed to flow down from the top of the glass tube, a substance having a strong adsorption force stops at the upper portion, and the weaker one flows first, so that the one having the weaker adsorption power flows out sequentially. Can be sorted. When an acidic substance derived from human sweat is used as a sample, for example, as shown in FIG. 5, a carboxylic acid containing a fatty acid having 12 or less carbon atoms other than β-hydroxycarboxylic acid is first used using a medium-polar solvent such as diethyl ether. After elution of the acids, the β-hydroxycarboxylic acid can be fractionated using a highly polar solvent such as methanol. Therefore, by adding a coloring reagent to the methanol fraction and coloring it, the presence of β-hydroxycarboxylic acid derived from human sweat can be confirmed. Further, the hue of the test liquid can be quantified not only by visual sensory evaluation but also by instrumental measurement using a spectrophotometer or a color difference meter.

  Thin-layer chromatography is a method of separating molecules having different polarities on a plate. For example, as shown in FIG. 6, after an acidic substance derived from human sweat is dropped at a predetermined position on a plate, diethyl acid is added to the plate. If developed with a medium polar solvent such as ether, fatty acids having 12 or less carbon atoms other than β-hydroxycarboxylic acid will spread around with the solvent, but β-hydroxycarboxylic acid will remain at the dropping position. Therefore, if a coloring reagent is dropped at the center of the dropping spot to give a color, the presence of sweat-derived β-hydroxycarboxylic acid can be confirmed. As described above, in the thin-layer chromatography method, since a coloring reagent can be directly sprayed on a plate, a color test for qualitative or quantitative determination can be rapidly performed.

  Methods for determining the presence or absence and the amount of β-hydroxycarboxylic acid using a color reaction include (1) a method of directly introducing a chromophore to a hydroxyl group or a carboxyl group of β-hydroxycarboxylic acid, and (2) ) A method of converting a β-hydroxycarboxylic acid into a derivative and then introducing a chromophore into the derivative; and (3) a method of decomposing the β-hydroxycarboxylic acid and introducing a chromophore into the decomposed product.

  (1) Coloring reagents that react with the carboxyl group of β-hydroxycarboxylic acid and develop color by use of a method for directly introducing a chromophore to the hydroxyl group or carboxyl group of β-hydroxycarboxylic acid include β-hydroxy A reagent for converting a carboxylic acid to a chromogenic acid hydrazide in the presence of a condensing agent to form a color, a reagent for converting a β-hydroxycarboxylic acid to a chromogenic ester to form a color, and a reagent for converting a β-hydroxycarboxylic acid to a chromogenic amide And color reagents.

  Coloring reagents leading to the chromogenic acid hydrazide include 2-nitrophenylhydrazine, 6,7-dimethoxy-1-methyl-2 (1H) -quinoxalinone-3-propionylcarboxylic acid hydrazide (DMEQ-H), p- (4,5-diphenyl-1H-imidazol-2-yl) -benzohydrazide, p- (1-methyl-1H-phenanthro- [9,10-d] imidazol-2-yl) -benzohydrazide, p- ( 5,6-dimethoxy-2-benzothiazoyl) -benzohydrazide and the like.

  Examples of reagents for leading a color-forming ester to form a color include 9-anthryldiazomethane, 1-naphthyldiazomethane, 1- (2-naphthyl) diazoethane, 1-pyrenyldiazomethane, 4-diazomethyl-7-methoxycoumarin, -Bromomethyl-7-methoxycoumarin, 3-bromomethyl-6,7-dimethoxy-1-methyl-2 (1H) -quinoxalinone, 9-bromomethylacridine, 4-bromomethyl-6,7-methylenedioxycoumarin, N- (9-acridinyl) -bromoacetamide, 2- (2,3-naphthylimino) ethyltrifluoromethanesulfonate, 2- (phthalimino) ethyltrifluoromethanesulfonate, N-chloromethylphthalimide, N-chloromethyl-4-nitrophthalimide, N -Chloromethyl isachi , O-(p-nitrobenzyl) -N, N'-diisopropyl isourea and the like.

  Examples of reagents that lead to a chromogenic amide to form a color include monodansyl cadaverine, 2- (p-aminomethylphenyl) -N, N'-dimethyl-2H-benzotriazol-5-amine, and the like.

  Coloring reagents that react with the hydroxyl group of β-hydroxycarboxylic acid to form a color include a reagent that leads to a chromogenic coordination compound to produce a color, a reagent that leads to a chromogenic ester to produce a color, and a coloring ether. And color reagents. Cerium ammonium nitrate and the like are examples of the reagent that leads to the color-forming coordination compound to form a color. Examples of the reagent that leads to a color-forming ester to form a color include 4- (2-phthalimidyl) benzoyl chloride, isomers thereof and derivatives thereof. Examples of the reagent that leads to a color-forming ether to form a color include 4-diazomethyl-7-methoxycoumarin and the like.

(2) In the method of converting a β-hydroxycarboxylic acid into a derivative and then introducing a chromophore into the derivative, as a β-hydroxycarboxylic acid derivative that can be used for a color reaction, an inorganic salt, hydroxamic acid, acid chloride, Examples thereof include a copper complex and a cobalt complex.
β-Hydroxycarboxylic acid inorganic salts react with aromatic halogens to form color-forming esters, hydroxamic acids to color-forming metal salts, acid chlorides to color-forming amides, copper complexes and cobalt complexes of hydroxycarboxylic acids. Respectively induces a chelate compound which is colored in response to copper or cobalt.

  As a method of converting β-hydroxycarboxylic acid into an inorganic salt, a method of neutralizing by mixing with an alkaline substance such as a sodium hydrogen carbonate solution, a sodium carbonate solution, a sodium hydroxide solution, and a potassium hydroxide solution may be mentioned. Examples of the aromatic halogen which can react with an inorganic salt of hydroxycarboxylic acid to form a color-forming ester include p-nitrobenzyl bromide, phenacyl bromide, p-chlorophenacyl bromide, p-bromophenacyl bromide, p-iodo. Phenacyl bromide, p-nitrophenacyl bromide, p-phenylphenacyl bromide, p-phenylazophenacyl bromide, N, N'-dimethyl-p-aminobenzeneazophenacyl chloride and the like can be mentioned.

  Examples of a method for converting β-hydroxycarboxylic acid to hydroxamic acid include a method of reacting with hydroxylamine in the presence of a condensing agent, and a method of reacting with hydroxylamine hydrochloride using nickel as a catalyst. Examples of the metal reagent which forms a complex having a color with hydroxamic acid include ferric chloride and vanadium.

  Examples of a method for converting β-hydroxycarboxylic acid to acid chloride include a method of reacting with oxalyl chloride. Examples of a method for converting acid chloride to a chromogenic amide include a method of reacting the acid chloride with 9-aminophenanthrene in the presence of triethylamine.

  Examples of the reagent that forms a color-forming chelate compound (complex) with copper or cobalt include diethyldithiocarbamic acid, bicyclohexanone oxalyldihydrazone, bathocuproine, and the like.

  (3) As a method for decomposing β-hydroxycarboxylic acid and coloring the decomposed product, β-hydroxycarboxylic acid is converted to acyl-amino acid in the presence of adenosine triphosphate (ATP) and coenzyme CoA. CoA synthetase acts to produce acyl-CoA, followed by treatment with acyl-CoA oxidase to produce enoyl-CoA and hydrogen peroxide, and further treatment of hydrogen peroxide with catalase to formaldehyde. And a colorimetric reagent, 4-amino-3-hydrazino-5-mercapto-1,2,3-triazole (AHMT), and reacting the resulting purple color.

  As described above, in the present invention, the reagent used for the color reaction of β-hydroxycarboxylic acid reacts with any of β-hydroxycarboxylic acid, β-hydroxycarboxylic acid derivative, and β-hydroxycarboxylic acid decomposition product to form a color. The compound having a hydrazino group such as 2-nitrophenylhydrazine (2-NPH), which is not particularly limited as long as it has high detection sensitivity to human sweat-derived β-hydroxycarboxylic acid and can be easily compared with the naked eye. Further, a compound having a diazomethyl group such as 9-anthryldiazomethane (ADAM), which has high detection sensitivity and allows the reaction to proceed under mild conditions, is preferable.

  The color reaction using 2-nitrophenylhydrazine is a reaction in which β-hydroxycarboxylic acid is reacted with 2-nitrophenylhydrazine in water or an alcohol solution in the presence of a condensing agent, and an acid which exhibits a purple-red color under alkaline conditions. This is a reaction for producing hydrazide (reaction formula (4)).

  As the condensing agent, dicyclohexylcarbodiimide (DCC) or 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) can be used. As the alkalizing agent, an aqueous solution of potassium hydroxide or the like can be used.

  When 0.5 mL of a water or alcohol solution containing 0.02 M of 2-nitrophenylhydrazine hydrochloride is added to 0.5 mL of a water or alcohol solution containing 0.01 to 0.05 μM of 3-hydroxy-3-methylhexanoic acid, The test solution has a slightly brownish yellow color. Further, when 0.5 mL of a water or alcohol solution containing 0.02 M of 2-nitrophenylhydrazine is added to 0.5 mL of an aqueous solution or an alcohol solution containing 0.05 to 1 μM of 3-hydroxy-3-methylhexanoic acid. The test solution develops a color from reddish brown to deep reddish purple. On the other hand, even if a water or alcohol solution containing 2-nitrophenylhydrazine is added to a water or alcohol solution containing no hydroxycarboxylic acid, no acid hydrazide is formed, and thus the color of the test solution is not measured. It remains unchanged from the yellow color derived from 2-nitrophenylhydrazine in the reaction.

  As described above, the intensity of apocrine odor is proportional to the content of β-hydroxycarboxylic acid in sweat, so by observing the degree of color development of the test solution with the naked eye, the degree of apocrine odor can be reliably, quickly, and rapidly increased. It can be easily determined.

  The color reaction using 9-anthryldiazomethane is a method for converting β-hydroxycarboxylic acid to 9-anthrylmethyl ester having strong fluorescence (reaction formula (5)).

  This reaction is completed at room temperature for about 10 to 60 minutes without the need for a catalyst or heating. 9-anthryl methyl ester has characteristics of an excitation wavelength of 365 nm and a fluorescence wavelength of 412 nm in methanol.

  After adding a 1.0% methanol or acetone solution of 9-anthryldiazomethane to a methanol solution containing 0.01 to 0.05 μM of 3-hydroxy-3-methylhexanoic acid, the mixture is slightly blue when illuminated with 365 nm light. After adding a 1.0% methanol or acetone solution of 9-anthryldiazomethane to a methanol solution containing 0.05 to 0.1 μM of 3-hydroxy-3-methylhexanoic acid, the solution was irradiated with light of 365 nm. When illuminated, the test solution emits strong blue fluorescence. On the other hand, if a 1.0% methanol-alcohol solution of 9-anthryldiazomethane is added to a solution containing no β-hydroxycarboxylic acid, no highly fluorescent ester is formed.

  As described above, the intensity of apocrine odor is proportional to the content of β-hydroxycarboxylic acid in sweat, so by observing the degree of color development of the test solution with the naked eye, the degree of apocrine odor can be reliably, quickly, and rapidly increased. It can be easily determined.

  In the present invention, when β-hydroxycarboxylic acid contained in sweat is isolated from axillary sweat and quantitatively evaluated with a coloring reagent, chemically synthesized 3-hydroxy-3-methylhexanoic acid is used as a standard substance. (Standard).

  In other words, β-hydroxycarboxylic acid contained in sweat can be more accurately quantified based on the degree of color development between the measured synthetic 3-hydroxy-3-methylhexanoic acid and the coloring reagent. At the same time, the amount of β-hydroxycarboxylic acid is proportional to the intensity of the apocrine odor, so that the degree of the apocrine odor can be accurately determined.

  The standard substance of 3-hydroxy-3-methylhexanoic acid used at that time may be used after being diluted to a considerable concentration in a laboratory, but in order to easily collect an amount suitable for comparison, methanol or acetone is used. It is more convenient to use it after diluting it in the above solvent. In particular, it is preferable to use, as a diluent of about 100 μL, 3-hydroxy-3-methylhexanoic acid in a range of 1 to 100 μg, and a standard solution diluted to an extent that can be measured.

  Further, in the present invention, when the difference in coloring between samples to be compared is delicate, or when determining how much the apocrine odor has been reduced after use of a deodorant or after apocrine sweat gland removal surgery, an analytical instrument By numerically expressing the hue of the test solution by using, the subjectivity can be excluded from the determination result.

  The analytical instrument used at this time is not particularly limited as long as it can measure the degree of color development of the test solution, but an acid hydrazide generated by a color reaction using 2-nitrophenylhydrazine includes a colorimeter and a colorimeter. An ultraviolet-visible spectrophotometer or the like can be used. In addition, a fluorescence spectrophotometer or the like can be used for 9-anthryl methyl ester produced by a color reaction using 9-anthryldiazomethane.

  Furthermore, in the present invention, not only the degree of apocrine odor that is specifically generated from the armpit of a person whose armpit is said to be constitutional, but also the acid odor generated from the armpit of a person regardless of the presence or absence of constitution (and also the sweat odor) ) Can also be determined quickly and easily.

  As a method of determining the degree of contribution of apocrine odor and acid odor, an acidic substance extracted from sweat is converted to β-hydroxycarboxylic acid causing apocrine odor and carbon number 12 or less in sweat causing acid odor. After separation into fatty acids, a color reagent is added to each of them, and a method of observing the developed color, or an acidic substance extracted from sweat and a β-hydroxycarboxylic acid separated from the acidic substance, respectively A method in which a color reagent is added thereto and the developed color is observed.

  As a method for determining the degree of acid odor, a coloring reagent may be added to a fatty acid having 12 or less carbon atoms other than β-hydroxycarboxylic acid in sweat which causes acid odor. For example, as shown in FIG. 5, in adsorption column chromatography, using diethyl ether as a developing solvent, eluting an acidic component derived from sweat, mainly a fatty acid having 12 or less carbon atoms other than β-hydroxycarboxylic acid, The β-hydroxycarboxylic acid was eluted using methanol, and a color reagent was added to the obtained ether fraction and methanol fraction, and the developed color was observed. The degree of contribution can be determined quickly and easily.

  In thin-layer chromatography, an acidic substance extracted from human sweat is dropped on a thin-layer chromatography plate, and a coloring reagent is dropped on the plate to cause the entire acidic substance to develop color. The color developed by the entire acidic substance is based on the total amount of β-hydroxycarboxylic acid, which is the main cause component of the apocrine odor, and the fatty acid having 12 or less carbon atoms other than β-hydroxycarboxylic acid, which is the main cause component of the acid odor. On the other hand, there is a positive correlation.

  On the other hand, the same acidic substance extracted was dropped into a similar thin-layer chromatoplate prepared separately, and β-hydroxycarboxylic acid and other fatty acids were separated with a developing solvent. -A color reagent is added dropwise to the hydroxycarboxylic acid to form a color. The color developed on this plate has a positive correlation with the amount of β-hydroxycarboxylic acid, which is a major cause of apocrine odor. Therefore, by observing the colors expressed in both, the degree of contribution of the apocrine odor and the acid odor can be determined.

  Therefore, by using the body odor determination kit according to the present invention, it is possible to quickly and easily determine the degree of contribution of apocrine odor and acid odor in each subject.

  The color reagent that reacts with the carboxylic acid contained in sweat to form a color is not particularly limited as long as it reacts with the carboxyl group of a fatty acid having 12 or less carbon atoms which greatly contributes to acid odor. The above-described color reagent that reacts with a carboxyl group of an acid to form a color can be used.

  In order to accurately determine the type and intensity of body odor, that is, the degree of contribution of each of the apocrine odor and the acid odor to sweat-derived acidic substances, the reactivity of the color reagent is determined by the β-hydroxycarboxylic acid and the β-hydroxycarboxylic acid. It is desirable that the specificity is high for fatty acids having 12 or less carbon atoms other than hydroxycarboxylic acid, and the color reaction to other acidic substances is weak.

  As described above, the body odor determination kit according to the present invention can measure the degree of the total armpit odor, which is a part of the body odor or a part of the body odor, without using an expensive analytical instrument such as gas chromatography or liquid chromatography. Since it is possible to make a reliable, quick and easy determination by taking into account the presence and strength of apocrine odor in the axillary area and also considering the presence and strength of acid odor, it can be used in hospitals (dermatologists, etc.) and health centers. It can be used to diagnose the degree of apocrine odor to a person who is interested in his or her own armpit odor, or to determine the effect of a deodorant agent or apocrine sweat gland removal surgery.

  In order to determine whether efforts to reduce body odor such as the use of a deodorant or apocrine sweat gland removal surgery are currently effective, for example, β-hydroxycarboxylic acid in sweat before and after use of a deodorant is used. And / or a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid may be quantitatively evaluated and compared using a main body odor determination kit. For example, if the sample was collected at a different time, the sweat sample collected first may be stored in an appropriate place such as a freezer so as not to cause a chemical change, and the determination test may be performed at the same time. When quantitatively evaluating the β-hydroxycarboxylic acid and fatty acids having 12 or less carbon atoms other than the β-hydroxycarboxylic acid in the test solution, if the color of the test solution is quantified by an analytical instrument, the timing of performing the judgment test differs. Even if they do, they can be accurately compared and evaluated.

  In addition, the kit for determining body odor according to the present invention can be used in a case of evaluating the effectiveness of a deodorant agent sample or in screening a person with a side arm in the development research of a deodorant agent targeting an apocrine odor. .

  As a method of determining the effectiveness of the deodorant agent targeting the apocrine odor, for example, a sample containing a predetermined concentration of β-hydroxycarboxylic acid, a predetermined amount of a deodorant agent sample is added, β-hydroxycarboxylic acid By evaluating the amount, preferably the amount of 3-hydroxy-3-methylhexanoic acid, using the body odor determination kit according to the present invention, the effectiveness of the deodorant sample can be determined accurately, quickly and easily.

  In addition, a deodorant sample is actually applied to the human axilla, and β-hydroxycarboxylic acid, preferably 3-hydroxy-3-methylhexanoic acid, contained in the axillary sweat collected before and after the application, is used in the present invention. It is also possible to evaluate and compare using the body odor judgment kit according to the above.

Specific examples of the configuration of the kit described above include, for example, the following, but are not limited thereto.
(1) A package in which absorbent cotton, a plastic cup, or the like for collecting human sweat is preferably processed into a usable state as it is, and is packaged with an exterior as needed.
(2) A package in which the components of the package described above are combined with an alkaline aqueous solution or an organic solvent for extracting an acidic substance from human sweat, a reagent such as a separating funnel, a solvent, and a device.
(3) Adsorption chromatography columns or thin-layer chromatography used to separate human sweat-derived β-hydroxycarboxylic acid and other fatty acids having 12 or less carbon atoms as components of any of the above packages. Package that combines instruments such as plates.
(4) A medium-polar solvent used for eluting fatty acids having 12 or less carbon atoms other than β-hydroxycarboxylic acid from acidic substances by adsorption or thin-layer chromatography, in combination with the components of any of the above packages. package.
(5) A package in which a component of any one of the above packages is combined with a highly polar solvent used for eluting β-hydroxycarboxylic acid from a column in adsorption chromatography.
(6) A package in which the components of any one of the above packages are filled with a coloring reagent that reacts with β-hydroxycarboxylic acid derived from human sweat, preferably in a state of being prepared in a composition or concentration usable as it is. A package that is combined and packaged as needed.
(7) A coloring reagent that reacts with a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid is preferably added to a component of any one of the above-mentioned packages to a composition or concentration that can be used as it is. A package that combines the contents of containers and packs them as needed.
(8) A coloring reagent that reacts with β-hydroxycarboxylic acid derived from human sweat and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid is preferably used as a component of any of the above packages, as it is. A package prepared by combining containers filled in a state where the composition or concentration has been adjusted to a possible level and packing the package as necessary.
(9) When the β-hydroxycarboxylic acid contained in sweat is isolated from the sweat in the axilla in any of the components of the package described above, when quantitatively evaluated with a color reagent, a preliminary preparation before the color reaction ( For example, for derivatization of β-hydroxycarboxylic acid).
(10) When the β-hydroxycarboxylic acid contained in the sweat is isolated from the sweat in the axillary region in any of the components of the package described above and then quantitatively evaluated using a color reagent, the degree of coloration of the test solution is determined. A package that combines equipment such as an ultraviolet lamp used for confirmation.
(11) using β-hydroxycarboxylic acid contained in sweat as a standard substance (standard) when quantitatively evaluating with a color reagent after isolating β-hydroxycarboxylic acid contained in sweat from constituents of any of the above packages; Preferably a package combining chemically synthesized 3-hydroxy-3-methylhexanoic acid or a diluent of 3-hydroxy-3-methylhexanoic acid.
(12) When the β-hydroxycarboxylic acid contained in sweat is isolated from the sweat in the axilla in any of the components of the package described above, when the color reagent is used for quantitative evaluation, the degree of color development of the test solution is determined. Package that combines a color sample sheet to perform.
(13) A package in which a dropping means such as a pipette is attached to a component of any one of the above-mentioned packages, which makes it easy to collect and drop a single use amount of the coloring reagent.
(14) A package in which analytical components such as a color difference meter and a spectrophotometer used to quantify the degree of coloration of a test solution are combined with the components of any of the above packages.

(Example 1)
Sixty-five healthy Japanese men were randomly selected as subjects. A new white T-shirt made of cotton was worn continuously for 24 hours. A sensory evaluation was performed on a portion of the T-shirt corresponding to the axilla by seven expert panelists. As a result, among the subjects, 52 people had strong acid odor, 10 people had strong apocrine odor, and 3 people felt both acid odor and apocrine odor.

(Example 2)
From the same group of subjects as in Example 1, three subjects having strong apocrine odor in the sensory evaluation were taken as subjects of Example 2.

  A cotton pad was sewn on a portion of the T-shirt that touches the axilla, the subject was allowed to wear the T-shirt for 24 hours, collected, and only acidic components were selectively extracted and concentrated from the axillary cotton pad according to a conventional method. The analysis was performed using a gas chromatography-mass spectrometer (GC-MS). The key components that generate the typical odor of axillary sweat were identified by sniffing GC.

  When the axillary odor component contained in the sweat was analyzed, GC-MS analysis (FIG. 1) revealed that together with the saturated fatty acids, 3-methyl-2-hexenoic acid, 7-octenoic acid, and γ-lactones, which had been conventionally confirmed, Showed the presence of 3-hydroxy-3-methylhexanoic acid. This eluted component had a strong odor very similar to the apocrine odor in smell gas chromatography. Further, it was found from the peak area that the compound was present at a high concentration.

  3-Hydroxy-3-methylhexanoic acid has never been reported as a component of human axillary odor, but was detected in three panels.

(Example 3)
From the same group of subjects as in Example 1, three subjects who had no apocrine odor in the sensory evaluation and were not recognized were regarded as subjects of Example 3. Axillary sweat was collected from this subject by the same procedure as in Example 2, and analyzed using GC-MS. As shown in FIG. 2, 3-hydroxy-3-methylhexanoic acid, which was observed in the group of subjects of Example 2, was not detected in any of the subjects.

(Example 4)
Among the same group of subjects as in Example 1, the group of subjects of Example 2 having a strong apocrine odor (three), the group of subjects of Example 3 having no apocrine odor (three), and the group of subjects having a mild apocrine odor For (2 persons), sweat in the axillary region was collected in the same procedure as in Example 2, and only the acidic component was selectively extracted according to a conventional method. After that, using a 1-ml volumetric flask (DURAN), an equal volume was used. Was prepared. This was analyzed using a gas chromatography-mass spectrometer (GC-MS) to quantify 3-hydroxy-3-methylhexanoic acid.
As shown in FIG. 3, as the apocrine odor increased, the amount of 3-hydroxy-3-methylhexanoic acid in the axilla increased.

(Example 5)
We asked 65 healthy Japanese men to volunteer. A new white T-shirt made of cotton was worn continuously for 24 hours. Seven professional panelists evaluated the apocrine odor intensity and the acid odor intensity of the portion corresponding to the axillary portion of the T-shirt according to the following criteria.
<Sensory evaluation criteria>
Strength 0 Does not smell.
１ 1 Smell slightly.
〃 2 Smell weakly.
〃 3 Smell.
４ 4 Smells a little stronger.
５ 5 Very strong smell.

  As a result, among the subjects, 10 persons with an intensity of 3 or more, 3 persons with an intensity of 1 or 2, and 52 persons with an intensity of 0 for the apocrine odor.

(Example 6)
Four people (Subject Nos. 1 to 4) with an apocrine odor of intensity 3 or more, three people (Subjects Nos. 5 to 7) with an apocrine odor of intensity 1 or 2, and an apocrine odor Three people (strength 0) who did not have this (subjects Nos. 8 to 10) were set as the subjects of Example 6.

  A cotton pad was sewn on a portion of the T-shirt corresponding to the axillary part on the dominant arm side, the subject was allowed to wear the T-shirt for 24 hours, collected, and acid-derived from sweat was removed from the cotton pad in the axillary part using an acid-base extraction method. The components were extracted. Next, the acidic extract was separated into a diethyl ether fraction and a methanol fraction using a silica gel mini column (Varian Bond Elute Jr), 10 mL of diethyl ether and 10 mL of methanol. After concentrating the methanol fraction once, a 1 mL diluted solution was prepared using a 1 mL volumetric flask (manufactured by DURAN).

  1 μL of the diluent was analyzed using a gas chromatography-mass spectrometer (GC-MS). Using the synthesized 3-hydroxy-3-methylhexanoic acid as a standard substance (standard), a calibration curve was prepared, and the amount of 3-hydroxy-3-methylhexanoic acid contained in sweat was measured.

  Table 1 shows the results of sensory evaluation of axillary odor and the results of quantification of 3-hydroxy-3-methylhexanoic acid. 3-Hydroxy-3-methylhexanoic acid was detected from the methanol fraction of all subjects (subjects Nos. 1 to 7) having an apocrine odor. On the other hand, it was not detected from the diethyl ether fraction.

  In the group in which an apocrine odor with an intensity of 3 or more was observed (subjects Nos. 1 to 4), the detected amount of 3-hydroxy-3-methylhexanoic acid was in the range of 15.71 to 64.29 μg. In the group in which the apocrine odor of intensity 1 or 2 was observed (subjects Nos. 5 to 7), the detected amount of 3-hydroxy-3-methylhexanoic acid was in the range of 0.87 to 1.71 µg. On the other hand, 3-hydroxy-3-methylhexanoic acid was not detected in any of the subjects in the group (subject Nos. 8 to 10) in which apocrine odor was not observed (strength 0).

(Example 7)
After concentrating the methanol fraction obtained in Example 6 (1 μL used for GC-MS analysis) to 0.5 mL, it was transferred to a test tube (manufactured by Maruem, NR-10) with a screw stopper. A blank sample for comparing hues (0.5 mL of methanol was added), 100 μL of a 20 mM 2-nitrophenylhydrazine solution prepared with 40 mM hydrochloric acid-ethanol hydrochloride (3: 1, v / v), and 3 v / v% pyridine 100 μL of an ethanol solution and 100 μL of an ethanol solution of 250 mM EDC (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide) were sequentially added. After heating this mixture at 60 ° C. for 20 minutes, 50 μL of a 15% (w / v) potassium hydroxide solution prepared with a water-methanol mixture (1: 4, v / v) was added. Heat at 60 ° C. until blank brown disappears (15 minutes). After cooling to room temperature, the color of the reaction solution was visually observed.

  Table 2 shows the results. The blank sample exhibited a bright yellow color derived from the unreacted reagent, but the methanol fraction of the subject in which 3-hydroxy-3-methylhexanoic acid was detected exhibited a purple-red color accompanying the production of acid hydrazide. The hue changed from slightly brownish yellow to deep reddish purple in proportion to the amount of detection.

  In the group in which the apocrine odor with an intensity of 3 or more was observed, the color of the reaction solution was in a range from red to reddish purple. In the group in which the apocrine odor of intensity 1 or 2 was observed, the color of the reaction solution was slightly brownish yellow. On the other hand, in the group in which no apocrine odor was observed (intensity 0), the test liquid of any subject was not colored, and had the same hue as the blank sample.

Next, the hue of the test liquid was digitized using a colorimeter. The color difference (ΔE ^* ab) is plotted on the horizontal axis, and the sensory evaluation result of the apocrine odor in Example 5 is plotted on the vertical axis. As shown in FIG. 7, the color difference (ΔE ^* ab) increased in proportion to the intensity of the apocrine odor.

<Measurement conditions by color difference meter>
Equipment used: Minolta colorimeter CT-310
Cell optical path length: 2mm
Temperature: 23 ° C
From the measurement data of the blank sample, the color difference was determined by the following equation, using the color difference reference colors (L ^* _t , a ^* _t , b ^* _t ) and the measurement data as (L ^* , a ^* , b ^* ).

(Example 8)
1 mL of the diethyl ether fraction obtained in Example 6 was transferred to a test tube with a screw stopper (manufactured by Maruem, NR-10). The solvent was distilled off on a water bath at 45 ° C., and 0.5 ml of methanol was added. Then, along with a blank sample for comparison of hue (0.5 ml of methanol was added), a coloring reagent was prepared in the same procedure as in Example 7. The color was developed using. The hue was quantified using a visible-ultraviolet analyzer together with the methanol fraction obtained in Example 7. The absorbance of the methanol fraction was plotted on the ordinate and the absorbance of the ether fraction was plotted on the abscissa, and the degree of contribution of apocrine odor and acid odor to each subject was evaluated.

  As shown in FIG. 8, the color of the methanol fraction of the acidic extract was strong, that is, the apocrine odor was observed, and the color (absorption) of the diethyl ether fraction was also strong. On the other hand, those who did not have an apocrine odor, that is, those whose methanol fraction did not develop color, tended to have weak coloration (absorption) of the diethyl ether fraction.

<Measurement conditions by UV-visible spectrometer>
Equipment used: BECHMAN DU-600
Cell optical path length: 10mm
Temperature: 23 ° C
Data measurement method: At 530 nm, the peak area of the spectrum obtained by subtracting the absorption spectrum of the blank sample from the absorption spectrum of the sample was measured.

(Example 9)
Ten subjects of Example 6 were set as subjects of Example 9. The methanol fraction of the acidic extract of sweat separated in the same manner as in Example 6 was concentrated to 0.5 ml, and then transferred to a test tube. 25 μL of a 1.0% acetone solution of 9-anthryldiazomethane was added together with a blank sample for comparison of hues (0.5 mL of methanol was added), sealed, and allowed to stand at room temperature for about 1 hour. Thereafter, using an ultraviolet lamp (Ultra Violet / Long wavelength 365 nm type), the generated fluorescence was visually observed. The results are shown in Table 3.

  In Table 3, the sample of the test subject in which the apocrine odor was recognized in Example 5 showed a result of emitting stronger fluorescence in proportion to the intensity of the apocrine odor. In contrast, no strong fluorescence was observed from the blank sample.

It is an elution peak which shows the result of having analyzed the sweat of the person who has an apocrine smell by GC-MS. It is an elution peak which shows the result of having analyzed the sweat of the person who does not have an apocrine smell by GC-MS. It is a graph which shows the relationship between the amount of 3-hydroxy-3-methylhexanoic acid contained in the sweat of the axilla, and the intensity of the apocrine odor. It is the figure which showed the procedure which judges the degree of body odor. It is a figure showing how to use a judgment kit at the time of judging a degree of body odor using an adsorption column chromatography method. It is a figure showing how to use a judgment kit at the time of judging a degree of body odor using thin layer chromatography. FIG. 4 is a diagram showing a relationship between apocrine odor intensity (sensory evaluation) and a color difference of a test solution after a color reaction. FIG. 3 is a view showing the absorbance of a methanol fraction and an ether fraction after a color reaction.

Claims (11)

An indicator substance for body odor determination containing at least one selected from the group consisting of β-hydroxy acid compounds represented by the following formula (1).

(In the formula, R ¹ is an alkyl having 1 to 4 carbons, R ² is a hydrogen atom or an alkyl having 1 to 4 carbons, and the total number of carbons in the formula (1) is 10 or less.) An indicator substance for body odor determination containing a β-hydroxy acid derivative obtained by introducing an atom or an atomic group into a β-hydroxyl group and / or a carboxyl group of a β-hydroxy acid compound represented by the following formula (1).

(In the formula, R ¹ is an alkyl having 1 to 4 carbons, R ² is a hydrogen atom or an alkyl having 1 to 4 carbons, and the total number of carbons in the formula (1) is 10 or less.)   A method for determining body odor using the indicator substance according to claim 1.   A method for determining the effectiveness of a deodorant agent using the indicator substance according to claim 1.   A human body odor determination kit comprising a color reagent that reacts with human sweat-derived β-hydroxycarboxylic acid.   The human body odor determination kit according to claim 5, further comprising a color reagent that reacts with a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid.   The determination kit according to claim 5 or 6, wherein the reagent comprises a compound having a hydrazino group or a diazomethyl group as an essential component.   The determination kit according to claim 7, wherein the reagent is 2-nitrophenylhydrazine or 9-anthryldiazomethane. A first step of extracting a mixture of β-hydroxycarboxylic acid contained in human sweat and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid,
A second step of adding the reagent to the mixture to form a color,
A third step of determining the type and / or intensity of the body odor according to the color developed in the second step. A first step of extracting a mixture of β-hydroxycarboxylic acid contained in human sweat and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid,
A second step of separating β-hydroxycarboxylic acid from the mixture;
A third step of reacting the reagent with the β-hydroxycarboxylic acid separated in the second step to form a color,
Determining the type and / or intensity of the body odor according to the color developed in the third step. A first step of extracting a mixture of β-hydroxycarboxylic acid contained in human sweat and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid,
A second step of separating the mixture into β-hydroxycarboxylic acid and a fatty acid having 12 or less carbon atoms other than the β-hydroxycarboxylic acid,
A third step of reacting the reagent with the β-hydroxycarboxylic acid separated in the second step to form a color,
A fourth step of reacting the reagent with a fatty acid having 12 or less carbon atoms other than β-hydroxycarboxylic acid separated in the second step to form a color,
A fifth step of determining the type and / or intensity of the body odor according to the colors developed in the third and fourth steps.

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