U.S. patent application number 10/529897 was filed with the patent office on 2006-07-20 for indicator for assessing body odor, process for producing the same, body odor assessment method, method of assessing efficaciousness of deodorant and kit for conveniently assessing body odor.
Invention is credited to Yoshihiro Hasegawa, Masamoto Matsukane, Emi Yabe.
Application Number | 20060160240 10/529897 |
Document ID | / |
Family ID | 32074730 |
Filed Date | 2006-07-20 |
United States Patent
Application |
20060160240 |
Kind Code |
A1 |
Hasegawa; Yoshihiro ; et
al. |
July 20, 2006 |
Indicator for assessing body odor, process for producing the same,
body odor assessment method, method of assessing efficaciousness of
deodorant and kit for conveniently assessing body odor
Abstract
An indicator material for assessing body odor of the present
invention comprises at least one member selected from the group
consisting of a substance (A) which is a .beta.-hydroxycarboxylic
acid compound represented by the formula (1), a substance (B) which
is a derivative of the compound represented by the formula (1), a
substance (C) which is an alcohol compound having a mercapto group
at the 3-position represented by the formula (2),and a substance
(D) which is a derivative of the compound represented by the
formula (2). The present invention also provides a method of
producing the compound represented by the formula (2) by incubating
perspiration originated from a human in an environment with an
oxygen concentration of 10 v/v % or less. The present invention
also provides a kit for assessing body odor of a human, wherein the
kit for assessing body odor of a human includes a coloration
reagent which reacts with .beta.-hydroxycarboxylic acid originated
from perspiration of a human.
Inventors: |
Hasegawa; Yoshihiro; (Tokyo,
JP) ; Matsukane; Masamoto; (Tokyo, JP) ; Yabe;
Emi; (Tokyo, JP) |
Correspondence
Address: |
OBLON, SPIVAK, MCCLELLAND, MAIER & NEUSTADT, P.C.
1940 DUKE STREET
ALEXANDRIA
VA
22314
US
|
Family ID: |
32074730 |
Appl. No.: |
10/529897 |
Filed: |
October 6, 2003 |
PCT Filed: |
October 6, 2003 |
PCT NO: |
PCT/JP03/12793 |
371 Date: |
December 16, 2005 |
Current U.S.
Class: |
436/166 ;
436/106 |
Current CPC
Class: |
Y10T 436/18 20150115;
Y10T 436/17 20150115; G01N 33/497 20130101; Y10T 436/201666
20150115; Y10T 436/182 20150115; C09B 26/02 20130101 |
Class at
Publication: |
436/166 ;
436/106 |
International
Class: |
G01N 33/00 20060101
G01N033/00 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 4, 2002 |
JP |
2002-293104 |
Mar 25, 2003 |
JP |
2003-083801 |
Apr 22, 2003 |
JP |
2003-116582 |
Jun 4, 2003 |
JP |
2003-160082 |
Oct 6, 2003 |
JP |
2003-346586 |
Claims
1. An indicator material for assessing body odor comprising at
least one member selected from the group consisting of: a substance
(A) which is a .beta.-hydroxycarboxylic acid compound represented
by the following formula (1): ##STR13## wherein R.sup.1 is an alkyl
having 1 to 4 carbons; R.sup.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 substance (B) which is a derivative of
.beta.-hydroxycarboxylic acid, wherein an atom(s) or an atomic
group(s) is introduced to a hydroxyl group and/or a carboxylic
group of a .beta.-hydroxycarboxylic acid compound represented by
the formula (1); a substance (C) which is an alcohol compound
having a mercapto group at the 3-position represented by the
following formula (2): ##STR14## wherein R.sup.3 is a hydrogen atom
or methyl group; R.sup.4 is an alkyl group having 1 to 3 carbons;
and R.sup.5 is a hydrogen atom or a methyl group, the total number
of carbons in the formula (2) is 8 or less; and a substance (D)
which is a derivative of an alcohol compound having a mercapto
group at the 3-position, wherein an atom(s) or an atom group(s) is
introduced to a mercapto group and/or a hydroxyl group of an
alcohol compound having a mercapto group at the 3-position
represented by the formula (2).
2. An indicator material for assessing body odor according to claim
1, wherein the indicator material for assessing body odor contains
the substance (A) and/or the substance (B).
3. An indicator material for assessing body odor according to claim
1, wherein the indicator material for assessing body odor contains
the substance (C) and/or the substance (D).
4. An indicator material for assessing body odor according to claim
1, wherein the indicator material for assessing body odor contains
the substance(s) (A) and/or (B) and the substance(s) (C) and/or
(D).
5. An indicator material for assessing body odor according to claim
4, wherein the indicator material for assessing body odor contains
the substance (A) and the substance (C).
6. An indicator material for assessing body odor according to claim
4, wherein the indicator material for assessing body odor contains
the substance (B) and the substance (D).
7. An indicator material for assessing body odor according to claim
5, wherein the weight ratio of the substances (C) and (A)
(substance (C):substance (A)) is 1:10 to 1:1,000.
8. A method of assessing body odor using as an index at least one
member selected from the group consisting of: a substance (A) which
is a .beta.-hydroxycarboxylic acid compound represented by the
following formula (1): ##STR15## wherein R.sup.1 is an alkyl having
1 to 4 carbons; R.sup.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 substance (B) which is a derivative of
.beta.-hydroxycarboxylic acid, wherein an atom(s) or an atomic
group(s) is introduced to a hydroxyl group and/or a carboxylic
group of a .beta.-hydroxycarboxylic acid compound represented by
the formula (1); a substance (C) which is an alcohol compound
having a mercapto group at the 3-position represented by the
following formula (2): ##STR16## wherein R.sup.3 is a hydrogen atom
or methyl group; R.sup.4 is an alkyl group having 1 to 3 carbons;
and R.sup.5 is a hydrogen atom or a methyl group, the total number
of carbons in the formula (2) is 8 or less; and a substance (D)
which is a derivative of an alcohol compound having a mercapto
group at the 3-position, wherein an atom(s) or an atom group(s) is
introduced to a mercapto group and/or a hydroxyl group of an
alcohol compound having a mercapto group at the 3-position
represented by the formula (2).
9. A method of assessing body odor according to claim 8, wherein an
indicator material comprising at least one member selected from the
group consisting of the substances (A), (B), (C) and (D) is
used.
10. A method of assessing body odor according to claim 8, wherein
the substance (A) and/or the substance (B) is used as an
index(es).
11. A method of assessing body odor according to claim 8, wherein
the substance (C) and/or the substance (D) is used as an
index(es).
12. A method of assessing body odor according to claim 8, wherein
the substances (A) and/or (B) and the substances (C) and/or (D) are
used as indexes.
13. A method of assessing body odor according to claim 12, wherein
the substance (A) and the substance (C) are used as indexes.
14. A method of assessing body odor according to claim 12, wherein
the substance (B) and the substance (D) are used as indexes.
15. A method of assessing body odor according to claim 13, wherein
an indicator material in which the weight ratio of the substances
(C) and (A) (substance (C):substance (A)) is 1:10 to 1: 1,000 is
used.
16. A method of assessing effectiveness of a deodorant using as an
index(es) at least one member selected from the group consisting
of: a substance (A) which is a .beta.-hydroxycarboxylic acid
compound represented by the following formula (1): ##STR17##
wherein R.sup.1 is an alkyl having 1 to 4 carbons; R.sup.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 substance (B)
which is a derivative of .beta.-hydroxycarboxylic acid, wherein an
atom(s) or an atomic group(s) is introduced to a hydroxyl group
and/or a carboxylic group of a .beta.-hydroxycarboxylic acid
compound represented by the formula (1); a substance (C) which is
an alcohol compound having a mercapto group at the 3-position
represented by the following formula (2): ##STR18## wherein R.sup.3
is a hydrogen atom or methyl group; R.sup.4 is an alkyl group
having 1 to 3 carbons; and R.sup.5 is a hydrogen atom or a methyl
group, the total number of carbons in the formula (2) is 8 or less;
and a substance (D) which is a derivative of an alcohol compound
having a mercapto group at the 3-position, wherein an atom(s) or an
atom group(s) is introduced to a mercapto group and/or a hydroxyl
group of an alcohol compound having a mercapto group at the
3-position represented by the formula (2).
17. A method of assessing effectiveness of a deodorant according to
claim 16, using an indicator material comprising at least one
member selected from the group consisting of the substances (A),
(B), (C) and (D).
18. AA method of assessing effectiveness of a deodorant according
to claim 16, using the substance (A) and/or the substance (B) as an
index(es).
19. AA method of assessing effectiveness of a deodorant according
to claim 16, using the substance (C) and/or the substance (D) as an
index(es).
20. AA method of assessing effectiveness of a deodorant according
to claim 16, using the substances (A) and/or (B) and the substances
(C) and/or (D) as indexes.
21. AA method of assessing effectiveness of a deodorant according
to claim 20, using the substance (A) and the substance (C) as
indexes.
22. AA method of assessing effectiveness of a deodorant according
to claim 20, using the substance (B) and the substance (D) as
indexes.
23. AA method of assessing effectiveness of a deodorant according
to claim 21, using an indicator material in which the weight ratio
of the substances (C) and (A) (substance (C):substance (A)) is 1:10
to 1:1,000.
24. AA method of producing an alcohol compound having a mercapto
group at the 3-position represented by the formula (2) comprising a
step of: incubating perspiration originated from a human in an
environment with an oxygen concentration of 10 v/v % or less:
##STR19## wherein R.sup.3 is a hydrogen atom or a methyl group;
R.sup.4 is an alkyl group having 1 to 3 carbons; and R.sup.5 is a
hydrogen atom or a methyl group, the total number of carbons in the
formula (2) is 8 or less.
25. A method of assessing body odor comprising steps of: incubating
perspiration originated from a human in an environment with an
oxygen concentration of 10 v/v % or less to produce an alcohol
compound having a mercapto group at the 3-position represented by
the formula (2); and using the produced compound as an index:
##STR20##
26. A method of assessing effectiveness of a deodorant comprising
steps of: incubating perspiration originated from a human in an
environment with an oxygen concentration of 10 v/v % or less to
produce an alcohol compound having a mercapto group at the
3-position represented by the formula (2); and using the produced
compound as an index: ##STR21##
27. A kit for assessing body odor of a human, wherein the kit for
assessing body odor of a human includes a coloration reagent which
reacts with .beta.-hydroxycarboxylic acid originated from
perspiration of a human.
28. A kit for assessing body odor of a human according to claim 27,
wherein the kit for assessing body odor of a human further includes
a coloration reagent which reacts with fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid.
29. A kit for assessing according to claim 27, wherein the reagent
includes a compound having a hydrazino group or a diazomethyl group
as an essential component.
30. A kit for assessing according to claim 29, wherein the reagent
is 2-nitrophenylhydrazine or 9-anthryldiazomethane.
31. A method of assessing body odor of a human comprising steps of:
a first step of extracting a mixture of .beta.-hydroxycarboxylic
acid and fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid from perspiration of a human; a
second step of adding the reagent to the mixture to exhibit color;
and a third step of assessing the kind and/or strength of body odor
from the color exhibited in the second step.
32. A method of assessing body odor of a human comprising steps of:
a first step of extracting a mixture of .beta.-hydroxycarboxylic
acid and fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid from perspiration of a human; a
second step of separating .beta.-hydroxycarboxylic acid from the
mixture; a third step of reacting said .beta.-hydroxycarboxylic
acid separated in the second step with the reagent to exhibit
color; and a fourth step of assessing the kind and/or strength of
body odor from the color exhibited in the third step.
33. A method of assessing body odor of a human comprising steps of:
a first step of extracting a mixture of .beta.-hydroxycarboxylic
acid and fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid from perspiration of a human; a
second step of separating the mixture into .beta.-hydroxycarboxylic
acid and fatty acid having 12 or less carbons other than said
0-hydroxycarboxylic acid respectively; a third step of reacting
said P-hydroxycarboxylic acid separated in the second step with the
reagent to exhibit color; a fourth step of reacting said fatty acid
having 12 or less carbons other than said .beta.-hydroxycarboxylic
acid separated in the second step with the reagent to exhibit
color; a fifth step of assessing the kind and/or strength of body
odor from each of the colors exhibited in the third and fourth
steps.
Description
TECHNICAL FIELD
[0001] The present invention relates to an indicator material for
assessing a body odor, and a method for assessing a level of a body
odor or effectiveness of a deodorant.
[0002] Also, the present invention relates to an assessing kit and
an assessment method including a coloration reagent, which reacts
.beta.-hydroxycarboxylic acid or the like originated from
perspiration of a human with the coloration reagent.
BACKGROUND ART
[0003] Recently, number of people who are anxious about their body
odor is increasing according to the growing desire for hygiene. The
body odor is a collective term for an odor generated from any parts
of a body, mainly a head, a mouth, underarms, a genital area, feet
or the like. Particularly, since an axillary odor (odor of
underarms) is easily sensed by a person oneself or people around, a
level thereof, for instance, some factors including the presence,
strength, difference in quality or the like of the axillary odor
tends to dominate the level of body odor in terms of a whole body.
Further, the person oneself or people around often feels the
axillary odor itself uncomfortable rather than the body odor.
[0004] In the axillary region of a human, there are not only
eccrine glands distributed almost all over the body but also
peculiar sweat glands called as apocrine glands. The axillary
region of a human is a part in which perspiration is less likely to
vaporize and bacteria are likely to grow. Hence, it is likely that
perspiration secreted from two kinds of sweat glands (eccrine sweat
and apocrine sweat), sebum, scurf or the like are metabolized by
bacteria on the skin to produce an odor.
[0005] An odor due to an eccrine sweat is called as a lower fatty
acid odor or simply an acid odor, which is caused by a lower
carboxylic acid having 2 to 5 carbons and smells sour and stuffy
(hereinafter called as a lower fatty acid odor or an acid odor).
Also, such an odor is not only produced in the underarm but also in
all skin surfaces of the whole body. On the other hand, an odor
derived from the apocrine sweat is produced in an axillary region
of a human having so called tragomaschalia habitus and called as an
apocrine odor or simply an "axillary odor" to distinguish. The
apocrine odor is a pungent odor peculiar to an axillary region and
particularly easily sensed by a person oneself or people around.
There are individual differences in actual axillary odors, which
can be roughly classified into an acid odor, an apocrine odor and
the mixed odor thereof.
[0006] The people of tragomaschalia habitus tend to have more
apocrine glands in the axillary region wherein an acid odor due to
an eccrine sweat and an apocrine odor are mixed to produce a strong
peculiar odor. Thus, if a person is anxious about own axillary
odor, the person tries to reduce the odor by using a deodorant
having a deodorant effect or germicidal effect, or by removing
apocrine glands of the axillary region in a surgical way if the
apocrine odor is particularly strong.
[0007] Hence, people who are anxious about own body odor or
axillary odor, people who have their body odor pointed out at home,
school office or the like by others, or people who may place their
underarms in front of someone's face such as hair stylists and
dentists even if their odors are in unnoticeable level by
themselves have great interest in how much apocrine odor they
originally have in their axillary regions, and further whether the
use of deodorants or the effort to reduce a body odor such as an
operation to remove apocrine glands is currently effective.
[0008] Conventionally, as methods of assessing a body odor,
particularly an apocrine odor, there are: (1) an organoleptic test
wherein a third party smells to determine an odor of perspiration
of axillary regions with one's nose; (2) an empirical assessing
method which presumes from the facts which are considered to be
relative to an apocrine odor such as genetic information, e.g.
whether there is a family member having tragomaschalia habit, a wet
cerumen, coloring of an underwear in the axillary regions or the
like; (3) a method which presumes from number and size of apocrine
glands, or the like.
[0009] The method (1) needs professional panel members such as a
skilled dermatologist, thus, it cannot be easily performed. Also,
the method (1) assesses the level of apocrine odor by smelling an
odor of the cotton wool which wiped underarms of a test subject,
which allows to mingle a judgment with a large degree of the
assessor's subjectivity and has difficulty in a quantitative
evaluation. Further, when the evaluation is performed
consecutively, a sense of smell may get tired so as to lower
objectivity.
[0010] The method (2) judges from genetic potential or indirectly
judges from a relationship between a wet cerumen and an apocrine
odor. The assessment by coloring of underwear focuses on coloring
matter contained in perspiration of apocrine glands, which does not
directly evaluate the apocrine odor of axillary regions, thus, it
is an indirect assessment method.
[0011] Therefore, there is a risk of false assessment with limited
evaluation points and there is a possibility of ignoring axillary
odor which is not actualized. Also, such studies can be rough
indications for assessing tragomaschalia habit, however, since they
are not quantitative and hard to assess, they may lack accuracy and
be not practical to assess the improvement in axillary odor after
the operation to remove apocrine glands, the presence and level of
recurrence of apocrine odor due to regeneration of apocrine glands
after the operation, the effectiveness of deodorizing or masking of
a deodorant or the like.
[0012] The method (3) estimates a level of a body odor in such
manner that a doctor makes a surgical incision in axillary regions
and then observes number and size of apocrine glands. A test
subject needs to accept mental and physical suffering, thus, the
method cannot be easily performed. Also, the operation fee is
generally high.
[0013] Trans-3-methyl-2-hexenoic acid, 7-octenoic acid or the like
contained in perspiration of underarms as odor components which are
distinctive of underarms is disclosed in "Molecular Recognition of
Taste and Smell," Kagaku Sosetsu No. 40, 205-211, (1999).
[0014] An use of certain .beta.-hydroxycarboxylic acid or the salt
thereof as an animal perfume material is disclosed in Japanese
Patent Application Laid-Open (JP-A) No. Hei. 10-25265.
[0015] On the other hand, a compound having a thiol group at the
3-position which has an effect to provide a significantly strong
clary sage-like odor as a flavoring component is disclosed in JP-A
No. 2000-95753.
[0016] 3-Mercapto-3-methyl-hexane-1-ol and
3-mercapto-2-methyl-butane-1-ol are disclosed as alcohol compounds
having a mercapto group at the 3-position in JP-A No. 2001-2634.
They are disclosed as flavoring components having characteristics,
wherein a S isomer of 3-mercapto-3-methyl-hexane-1-ol has a
grass-like or agrestic odor, a R isomer of
3-mercapto-3-methyl-hexane-1-ol has a grapefruit/passion
fruit-like, currant-like or onion-like odor, and
3-mercapto-2-methyl-butane-1-ol has a grass-like, leek-like and
gas-like odor.
[0017] 3-Mercapto alcohol and formic and acetic esters thereof are
disclosed in German Patent Application Laid-Open No. 2,316,456 as
fragrance agents and flavor agents effective for preparation and
modification of a wide range of flavor components, wherein said
3-mercapto alcohol and formic and acetic esters thereof have
onion-like, sulfur-like or sweat-like odor.
[0018] A thio derivative used as a flavor component and/or a flavor
enhancer is disclosed in JP-A No. 2003-12637, wherein the thio
derivative reminds of smell of a black currant, onion or
grapefruit.
[0019] However, none of .beta.-hydroxycarboxylic acid and/or
derivative thereof and the alcohol compounds having a mercapto
group at the 3-position and/or derivatives thereof was recognized
to have a relationship with a body odor of a human particularly in
terms of a causative agent of a body odor or a constituent of an
axillary odor of a human. Hence, presence or strength of a body
odor of a human, particularly, an apocrine odor of axillary regions
has not been able to be assessed objectively and
quantitatively.
[0020] Also, as a characteristic component of the apocrine odor in
axillary regions of a human, there were conventionally not only
unsaturated carboxylic acid having 6 to 10 carbons as typified by
trans-3-methyl-2-hexenoic acid and 7-octenoic acid but also lower
carboxylic acid having 5 or less carbons which causes acid odor and
higher carboxylic acid originated from sebum in mixture. It has
been difficult to selectively separate the unsaturated carboxylic
acid which is a characteristic component of an apocrine odor among
the various acid components
DISCLOSURE OF INVENTION
[0021] An object of the present invention is to provide an
indicator material for assessing body odor comprising at least one
member selected from the group consisting of:
[0022] a substance (A) which is a .beta.-hydroxycarboxylic acid
compound represented by the following formula (1): ##STR1## wherein
R.sup.1 is an alkyl having 1 to 4 carbons; R.sup.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;
[0023] a substance (B) which is a derivative of
.beta.-hydroxycarboxylic acid, wherein an atom(s) or an atomic
group(s) is introduced to a hydroxyl group and/or a carboxylic
group of a .beta.-hydroxycarboxylic acid compound represented by
the formula (1);
[0024] a substance (C) which is an alcohol compound having a
mercapto group at the 3-position represented by the following
formula (2): ##STR2## wherein R.sup.3is a hydrogen atom or methyl
group; R.sup.4 is an alkyl group having 1 to 3 carbons; and R.sup.5
is a hydrogen atom or a methyl group, the total number of carbons
in the formula (2) is 8 or less; and
[0025] a substance (D) which is a derivative of an alcohol compound
having a mercapto group at the 3-position, wherein an atom(s) or an
atom group(s) is introduced to a mercapto group and/or a hydroxyl
group of an alcohol compound having a mercapto group at the
3-position represented by the formula (2).
DETAILED DESCRIPTION OF THE INVENTION
[0026] The present invention provides an indicator material capable
of assessing presence and strength of an apocrine odor of axillary
regions about which many people are particularly anxious among body
odors objectively and quantitatively, and a method using the
indicator material to assess a level of a body odor or
effectiveness of a deodorant. Also, the present invention provides
a method of producing the indicator material for a simple and
accurate assessment of a body odor. Further, the present invention
provides a kit capable of surely, promptly and easily assessing the
kind and level of body odor of a human, and a method of assessing
human odor using the kit.
[0027] As the result of diligent researches on components which
cause the apocrine odor contained in perspiration of underarms, the
inventors found out that substances exist in perspiration which
have a significantly similar odor to the apocrine odor and
characteristically exist in perspiration of people having the
apocrine odor, have enough concentration to accurately determine
the quantity as they are or by incubation, and can be separated by
a simple chemical operation. Further, the inventors made the
substances capable of being used as objective indexes for
quantitatively assessing a level of the apocrine odor of axillary
regions.
[0028] Also, various marker materials can be introduced to such
substances in order to increase analytical sensitivity and/or
accuracy of assessment, and obtained derivatives of indicator
materials can be suitably used for assessing body odor and
effectiveness of a deodorant.
[0029] According to the present invention, an indicator material
for assessing body odor is comprised of at least one member
selected from the group consisting of:
[0030] a substance (A) which is a .beta.-hydroxycarboxylic acid
compound represented by the following formula (1): ##STR3## wherein
R.sup.1 is an alkyl having 1 to 4 carbons; R.sup.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;
[0031] a substance (B) which is a derivative of
.beta.-hydroxycarboxylic acid, wherein an atom(s) or an atomic
group(s) is introduced to a hydroxyl group and/or a carboxylic
group of a .beta.-hydroxycarboxylic acid compound represented by
the formula (1);
[0032] a substance (C) which is an alcohol compound having a
mercapto group at the 3-position represented by the following
formula (2): ##STR4## wherein R.sup.3is a hydrogen atom or methyl
group; R.sup.4 is an alkyl group having 1 to 3 carbons; and
R.sup.5is a hydrogen atom or a methyl group, provided that the
total number of carbons in the formula (2) is 8 or less; and
[0033] a substance (D) which is a derivative of an alcohol compound
having a mercapto group at the 3-position, wherein an atom(s) or an
atom group(s) is introduced to a mercapto group and/or a hydroxyl
group of an alcohol compound having a mercapto group at the
3-position represented by the formula (2).
[0034] Also, a method of assessing body odor and a method of
assessing effectiveness of a deodorant according to the present
invention are methods of assessing the level of body odor or
effectiveness of a deodorant using as an indicator material at
least one member selected from the group consisting of the
above-mentioned substances (A), (B), (C) and (D). In the case of
selecting two or more indicator substances among the substances
(A), (B), (C) and (D) to use, an indicator material containing all
selected substances may be used or each of the selected indicator
substances may be used separately in the course of the assessment
process. Also, in a set of assessment process, a part of selected
indicator substances may be mixed to use and each of the substances
which is not mixed may be used separately as a sole substance.
[0035] In the present invention, one kind among the above-mentioned
substances (A), (B), (C), and (D) may be selected to use, or two or
more kinds of indicator substances may be arbitrarily selected to
use in combination. For example, the indicator substances may be
used singly or in combination as follows.
[0036] (1) An indicator substance comprising at least one member
selected from the group consisting of the substance (A) and/or the
substance (B).
[0037] The substance (A), i.e. a .beta.-hydroxycarboxylic acid
compound represented by the formula (1), is particularly effective
to evaluate a cumin oil-like apocrine odor. The substance (B) is a
derivative of the substance (A) and suitably used in lieu of the
substance (A) in order to increase analytical sensitivity and/or
accuracy of assessment, wherein the substance (B) may be used in
combination with the substance (A).
[0038] (2) An indicator substance comprising at least one member
selected from the group consisting of the substance (C) and/or the
substance (D).
[0039] The substance (C), i.e. an alcohol compound having a
mercapto group at the 3-position represented by the formula (2), is
particularly effective to evaluate a sulfur-like apocrine odor. The
substance (D) is a derivative of the substance (C) and suitably
used in lieu of the substance (C) in order to increase analytical
sensitivity and/or accuracy of assessment, wherein the substance
(D) may be used in combination with the substance (C).
[0040] (3) A combination of an indicator substance comprising at
least one member selected from the group consisting of the
substance (A) and/or the substance (B) and an indicator substance
comprising at least one member selected from the group consisting
of the substance (C) and/or the substance (D).
[0041] By using the indicator substance comprising the group
consisting of the substance (A) and/or the substance (B) in
combination with the indicator substance comprising the group
consisting of the substance (C) and/or the substance (D), both
cumin oil-like apocrine odor and sulfur-like apocrine odor can be
evaluated, thus, an accurate evaluation can be performed in
accordance with an actual apocrine odor.
[0042] (4) A combination of an indicator substance comprising at
least one member selected from the group consisting of the
substance (A) and an indicator substance comprising at least one
member selected from the group consisting of the substance (C).
[0043] This is a combination of indicator substances which are not
derivatized, which may be more suitable than using derivatives
depending on a method of analysis or evaluation.
[0044] (5) A combination of an indicator substance comprising at
least one member selected from the group consisting of the
substance (B) and an indicator substance comprising at least one
member selected from the group consisting of the substance (D).
[0045] This is a combination of indicator substances which are
derivatized, which can analyze or evaluate a level of odor by
methods other than a direct assessment using an organoleptic test
and also can enhance analytical sensitivity and/or accuracy of
assessment.
[0046] Moreover, a method of producing an indicator material
according to the present invention is a method of producing an
alcohol compound having a mercapto group at the 3-position
represented by the formula (2) by incubating the perspiration
originated from a human in an environment with 10 v/v % or less of
an oxygen concentration.
[0047] The perspiration originated from a human is incubated in an
environment with anaerobic or microaerophilic atmosphere to produce
an alcohol compound having a mercapto group at the 3-position in
large quantity, thereby, it is possible to make the assessment of
body odor and effectiveness of a deodorant easier and more
accurate.
[0048] Further, the present inventors have focused attention on a
point that a level of apocrine odor can be easily assessed from the
color exhibited by separating .beta.-hydroxycarboxylic acid
including 3-hydroxy-3-methylhexanoic acid which is newly found out
as a main component which causes apocrine odor from perspiration of
axillary regions, and thereafter reacting it with a coloration
reagent, and developed a kit capable of surely and easily assessing
a level of apocrine odor itself or a total body odor with focus on
apocrine odor, and a method of assessing human odor using the
kit.
[0049] The kit for assessing according to the present invention is
a kit for assessing body odor of a human including a coloration
reagent which reacts with .beta.-hydroxycarboxylic acid and/or
fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid originated from perspiration of a
human respectively. The kit enabled to assess a kind and strength
of odor and a level of human odor originated from perspiration of a
human surely, promptly and easily from the color exhibited by
reacting .beta.-hydroxycarboxylic acid which is a material causing
an apocrine odor and fatty acid having 12 or less carbons other
than said .beta.-hydroxycarboxylic acid which is a material causing
an acid odor with a coloration reagent.
[0050] A first method of assessing body odor using the kit of the
present invention is a method of assessing body odor of a human
comprising steps of:
[0051] a first step of extracting a mixture of
.beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid from
perspiration of a human;
[0052] a second step of adding the reagent to the mixture to
exhibit color; and
[0053] a third step of assessing the kind and/or strength of body
odor from the color exhibited in the second step.
[0054] Also, a second method of assessing body odor using the kit
of the present invention is a method of assessing body odor of a
human comprising steps of:
[0055] a first step of extracting a mixture of
.beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid from
perspiration of a human;
[0056] a second step of separating .beta.-hydroxycarboxylic acid
from the mixture;
[0057] a third step of reacting said .beta.-hydroxycarboxylic acid
separated in the second step with the reagent to exhibit color;
and
[0058] a fourth step of assessing the kind and/or strength of body
odor from the color exhibited in the third step.
[0059] Further, a third method of assessing body odor using the kit
of the present invention is a method of assessing body odor of a
human comprising steps of:
[0060] a first step of extracting a mixture of
.beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid from
perspiration of a human;
[0061] a second step of separating the mixture into
.beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid
respectively;
[0062] a third step of reacting said .beta.-hydroxycarboxylic acid
separated in the second step with the reagent to exhibit color;
[0063] a fourth step of reacting said fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid separated in
the second step with the reagent to exhibit color;
[0064] a fifth step of assessing the kind and/or strength of body
odor from each of the colors exhibited in the third and fourth
steps.
BRIEF DESCRIPTION OF DRAWINGS
[0065] In the accompanying drawings,
[0066] FIG. 1 shows the elution peaks as a result of GC-MS analysis
of an acid extract of perspiration of a person having an apocrine
odor;
[0067] FIG. 2 shows the elution peaks as a result of GC-MS analysis
of an acid extract of perspiration of a person having no apocrine
odor;
[0068] FIG. 3 is a graph showing the relationship between the
amounts of 3-hydroxy-3-methylhexanoic acid contained in
perspiration of axillary regions and the strength of an apocrine
odor;
[0069] FIG. 4 shows the result of GC-MS analysis of the
perspiration of a person having an apocrine odor;
[0070] FIG. 5 shows the result of GC-MS analysis of the
perspiration of a person not having apocrine odor;
[0071] FIG. 6 shows the result of GC-MS analysis of the
perspiration of a person having apocrine odor after incubation;
[0072] FIG. 7 shows the result of GC-MS analysis of the
perspiration of a person not having apocrine odor after
incubation;
[0073] FIG. 8 shows the process of assessing the level of body
odor;
[0074] FIG. 9 shows the method of using a kit for assessing the
level of body odor with the use of an absorption column
chromatography;
[0075] FIG. 10 shows the method of using a kit for assessing the
level of body odor with the use of a thin-layer column
chromatography;
[0076] FIG. 11 shows the relationship between the strength of
apocrine odor (organoleptic test) and the color differences of test
solutions after color reactions; and
[0077] FIG. 12 shows the absorbance of methanol fraction and ether
fraction after color reactions.
[0078] Hereinafter, the present invention will be explained in
detail. Contents of all publications cited in the present
specification are incorporated herein by reference.
[0079] According to the present invention, an indicator material
for assessing body odor which is quite similar to actual apocrine
odor is prepared by selecting one or more substances among a group
of .beta.-hydroxycarboxylic acid compounds including
3-hydroxy-3-methylhexanoic acid and compounds having a chemical
structure quite similar thereto, or selecting one or more
substances among a group of 3-mercapto alcohol compounds including
3-mercapto-3-methylhexanol and compounds having a chemical
structure quite similar thereto, or preferably by using the
.beta.-hydroxycarboxylic acid compound and the 3-mercapto alcohol
compound in combination, and thus an accurate evaluation result
with regard to the level (strength and quality) of body odor which
people actually feel and effectiveness of a deodorant against body
odor can be obtained.
[0080] It is considered that said 3-hydroxy-3-methylhexanoic acid
and 3-mercapto alcohol compounds found in perspiration of axillary
regions of a human by the inventors of the present invention have
the following features, thus, the presence amount and the presence
state of 3-hydroxy-3-methylhexanoic acid and/or 3-mercapto alcohol
compounds in axillary regions form the level and the individual
difference of apocrine odor.
(Features of 3-hydroxy-3-methylhexanoic acid and 3-mercapto alcohol
compound)
[0081] (1) A person that 3-hydroxy-3-methylhexanoic acid and/or
3-mercapto alcohol compound is not detected in the perspiration of
his axillary regions does not have the apocrine odor, but a person
that the 3-hydroxy-3-methylhexanoic acid and/or 3-mercapto alcohol
compound is detected in his perspiration of the axillary regions
has the apocrine odor. That is, the 3-hydroxy-3-methylhexanoic acid
and 3-mercapto alcohol compound are specifically present in the
person who has the apocrine odor (FIGS. 1, 2, 4, and 5).
[0082] (2) The apocrine odor is stronger in a person who has more
3-hydroxy-3-methylhexanoic acid (FIG. 3) and/or 3-mercapto alcohol
compound contained in the perspiration of the axillary regions.
[0083] (3) A person that 3-hydroxy-3-methylhexanoic acid and/or
3-mercapto alcohol compound is not detected in the perspiration of
his axillary regions even after incubation does not have the
apocrine odor. On the other hand, as for a person having the
apocrine odor, the amount of 3-hydroxy-3-methylhexanoic acid and/or
the 3-mercapto alcohol compound contained in the perspiration of
his axillary regions increases by incubation. That is,
3-hydroxy-3-methylhexanoic acid and the 3-mercapto alcohol compound
specifically increase by incubating the perspiration of the person
having the apocrine odor (FIGS. 6 and 7).
[0084] (4) As the .beta.-hydroxycarboxylic acid compound has not
only a carboxyl group but also a hydroxyl group at the 3-position,
a chemical modification can be performed in order to have good
detection sensitivity in instrumental analyses such as a gas
chromatography, a liquid chromatography or the like or to utilize
the coloration reaction for assessing by a spectrometer or naked
eye. Also, the .beta.-hydroxycarboxylic acid and/or the derivative
thereof can be separated from other substances which are low in
contribution to the apocrine odor utilizing differences in
polarity, solubility or the like.
[0085] (5) As the 3-mercapto alcohol compound has not only a
hydroxyl group but also a mercapto group at the 3-position, a
chemical modification can be performed in order to have good
detection sensitivity in instrumental analyses such as a gas
chromatography, a liquid chromatography or the like or to utilize
the coloration reaction for assessing by a spectrometer or naked
eye. Also, the 3-mercapto alcohol compound and/or the derivative
thereof can be separated from other substances which are low in
contribution to the apocrine odor utilizing differences in
polarity, solubility or the like.
[0086] (6) As the .beta.-hydroxycarboxylic acid compound and the
3-mercapto alcohol compound can be isolated by the adsorption
chromatography or the like, the color reaction, which is sure,
prompt and easy, can be utilized to evaluate quantitatively.
[0087] (7) Also, there are a spicy cumin-like odor and a fishy
sulfur-like odor as major bad odors which comprise the apocrine
odor. 3-Hydroxy-3-methylhexanoic acid is detected relatively in
abundance from a person who has the strong spicy cumin-like odor.
The 3-mercapto alcohol compound is detected relatively in abundance
from a person who has the fishy sulfur-like odor.
[0088] Moreover, the .beta.-hydroxycarboxylic acid compound, which
is a group of compounds having a chemical structure quite similar
to 3-hydroxy-3-methylhexanoic acid, is similar in characteristics
such as chemical characteristics or organoleptic characteristics
(particularly, odor) to 3-hydroxy-3-methylhexanoic acid, thus, the
.beta.-hydroxycarboxylic acid compound can be used as an objective
index for assessing apocrine odor similarly to
3-hydroxy-3-methylhexanoic acid.
[0089] Therefore, by measuring the presence amount and the presence
state of the .beta.-hydroxycarboxylic acid and/or the derivative
thereof (for instance, salt or ester of 3-hydroxy-3-methylhexanoic
acid) in axillary regions by an appropriate means which is
chemical, physical or the like, the apocrine odor in the axillary
regions can be measured objectively and quantitatively with the use
of the .beta.-hydroxycarboxylic acid compound and/or the derivative
thereof as an index.
[0090] Also, by measuring the presence amount and the presence
state of the 3-mercapto alcohol compound and/or the derivative
thereof (for instance, salt or ester of 3-mercapto-3-methylhexanol)
in axillary regions by an appropriate means which is chemical,
physical or the like, the apocrine odor in the axillary regions can
be measured objectively and quantitatively with the use of the
3-mercapto alcohol compound and/or the derivative thereof as an
index.
[0091] Particularly in the present invention, by using the
.beta.-hydroxycarboxylic acid compound and/or the derivative
thereof and the 3-mercapto alcohol compound and/or the derivative
thereof in combination, both cumin-like apocrine odor and
sulfur-like apocrine odor can be evaluated, thus, an accurate
evaluation can be performed in accordance with an actual apocrine
odor. Also, in the case of using such a combination for the
assessment of body odor and effectiveness of a deodorant, the level
of body odor of a human or axillary odor, which is a part of body
odor, can be assessed objectively and quantitatively not only from
presence and strength of apocrine odor in the axillary regions but
also from difference in quality thereof so that the accuracy of the
assessment result can be raised.
[0092] The .beta.-hydroxycarboxylic acid compound of the substance
(A) is a group of compounds including 3-hydroxy-3-methylhexanoic
acid and compounds having a chemical structure quite similar
thereto, which have an odor quite similar to the apocrine odor and
is represented by the following formula (1).
3-Hydroxy-3-methylhexanoic acid is represented by the following
formula (3): ##STR5## wherein R.sup.1 is an alkyl having 1 to 4
carbons; R.sup.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; ##STR6##
[0093] In the above formula (1), R.sup.1 is an alkyl having 1 to 4
carbons, which may be a straight or branched alkyl. For example,
there may be methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl
and t-butyl. It can be considered that the .beta.-hydroxycarboxylic
acid compound is easier to use as an indicator material if the
characteristic is closer to 3-hydroxy-3-methylhexanoic acid. Hence,
in order to have the chemical structure close to
3-hydroxy-3-methylhexanoic acid, it is preferable that R.sup.1 has
3 or 4 carbons, particularly the case of 3 carbons is preferable,
and is preferably a straight-chain alkyl.
[0094] In the above formula (1), R.sup.2 is a hydrogen atom or an
alkyl having 1 to 4 carbons, which may be a straight or branched
alkyl. As R.sup.2, for example, there may be methyl, ethyl,
n-propyl, i-propyl, n-butyl, i-butyl and t-butyl. In order to have
the chemical structure of the .beta.-hydroxycarboxylic acid
compound close to 3-hydroxy-3-methylhexanoic acid, it is preferable
that R.sup.2 has 1 or 2 carbons, particularly the case of 1 carbon
is preferable.
[0095] Among the .beta.-hydroxycarboxylic acid,
3-hydroxy-3-methylhexanoic acid, 3-hydroxy-3-methylpentanoic acid,
3-hydroxy-3-methylbutanoic acid, 3-hydroxyhexanoic acid and
3-hydroxypentanoic acid are preferable. Among the above,
3-hydroxy-3-methylhexanoic acid is particularly suitable as an
indicator material since 3-hydroxy-3-methylhexanoic acid itself is
a major causative substance of apocrine odor present in the
perspiration of the axillary regions as aforementioned.
[0096] 3-Mercapto alcohol compound of the substance (C) in the
present invention is contained relatively in large quantity in the
perspiration of a person having the apocrine odor, and it is a
group of compounds including 3-mercapto-3-methylhexanol,
3-mercaptohexanol, 3-mercaptopentanol, 3-mercapto-2-methylpentanol,
3-mercapto-2-methylbutanol and compounds having a chemical
structure quite similar to those compounds, which has odor quite
similar to the apocrine odor and is represented by the following
formula (2). 3-Mercapto-3-methylhexanol, 3-mercaptohexanol,
3-mercaptopentanol, 3-mercapto-2-methylbutanol and
3-mercapto-2-methylpentanol are represented by the following
formulae (4a) to (4e) ##STR7## wherein R.sup.3is a hydrogen atom or
methyl group; R.sup.4is an alkyl group having 1 to 3 carbons; and
R.sup.5 is a hydrogen atom or a methyl group, the total number of
carbons in the formula (2) is 8 or less. ##STR8##
[0097] It can be considered that the 3-mercapto alcohol compound is
easier to use as an indicator material if the characteristic is
closer to 3-mercapto-3-methylhexanol which is contained in the
perspiration in relatively large quantity among those compounds.
Hence, it is preferable to have the chemical structure close to
3-mercapto-3-methylhexanol.
[0098] From the viewpoint, R.sup.3 in the above formula (2) is
preferably methyl group among hydrogen atom and methyl group.
R.sup.4 is an alkyl group having 1 to 3 carbons, which may be a
straight or branched alkyl, and examples of which include methyl,
ethyl, n-propyl and i-propyl. Particularly, it is preferable that
R.sup.4 has 2 or 3 carbons, and more preferably 3 carbons. Also,
R.sup.5 may be a hydrogen atom or a methyl group. Among them, a
hydrogen atom is preferable.
[0099] Among the 3-mercapto alcohol compounds,
3-mercapto-3-methylhexanol, 3-mercaptohexanol, 3-mercaptopentanol,
3-mercapto-2-methylpentanol and 3-mercapto-2-methylbutanol are
preferable. Among them, 3-mercapto-3-methylhexanol is particularly
suitable for an indicator material since 3-mercapto-3-methylhexanol
is contained in the perspiration of the axillary regions relatively
in large quantity.
[0100] The substance (A) of the present invention may be subject to
chemical modification so as to be used as the substance (B) in
order to be detected in high sensitivity in the gas chromatography
or liquid chromatography or be assessed by a spectrometer or naked
eye utilizing the coloring reaction unless it does not lose the
detection function as an indicator material. For example, an
atom(s) or an atomic group(s) may be introduced to one or both of a
carboxyl group and/or a hydroxyl group in the .beta.-position of
the .beta.-hydroxycarboxylic acid compound and then used as
derivatives such as salt, ester, amide, ether or the like.
[0101] As a reagent which can be used in the liquid chromatography
analysis, spectrophotometer analysis or colorimetry test, and
reacts with a carboxyl group of the .beta.-hydroxycarboxylic acid
compound, there may be the reagents which can lead to acid
hydrazide such as 2-nitrophenylhydrazine,
6,7-dimethoxy-1-methyl-2(1H)-quinoxaline-3-propionyl carboxylic
acid hydrazide (DMEQ-H),
p-(4,5-diphenyl-1H-imidazole-2-il)-benzohydrazide,
p-(1-methyl-1H-phenanthro-[9,10-d]imidazole-2-il)-benzo hydrazide,
p-(5,6-dimethoxy-2-benzothiazoyl)-benzohydrazide or the like; the
reagents which can lead to ester such as 9-anthryldiazomethane,
1-naphthyldiazomethane, 1-(2-naphthyl)diazoethane,
1-pyrenyldiazomethane, 4-diazomethyl-7-methoxycoumarin,
4-bromomethyl-7-methoxycoumarin (Br-MmC),
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)ethyltrifluoromethane sulfonate,
2-(phthalimino)ethyltrifluoromethane sulfonate,
N-chloromethylphthalimide, N-chloromethyl-4-nitrophthalimide,
N-chloromethylisatin, o- (p-nitrobenzyl)-N,N'-diisopropylisourea
(PNBDI) or the like; the reagents which can lead to amide such as
monodansylcadaverine,
2-(p-aminomethylphenyl)-N,N-dimethyl-2H-benzotriazole-5-amine or
the like.
[0102] As a reagent which can be used in the liquid chromatography
analysis, spectrophotometer analysis and colorimetry test, and
reacts with a hydroxyl group of the .beta.-hydroxycarboxylic acid
compound, there may be the reagents such as cerium ammonium nitrate
in order to lead to a coordination compound, the reagents such as
4-(2-phthalimidyl) benzoyl chloride and the derivative thereof in
order to lead to ester, the reagents such as
4-diazomethyl-7-methoxycoumarin in order to lead to ester, or the
like.
[0103] Also, the .beta.-hydroxycarboxylic acid compound may be led
to inorganic salt, hydroxamic acid, acid chloride, a copper
complex, cobalt complex or the like, and then further led to a
chromophiric compound. The inorganic salt of the
.beta.-hydroxycarboxylic acid compound may be led to a chromophiric
ester, the hydroxamic acid may be led to a chromophiric metal salt,
acid chloride may be led to a chromophiric amide, and a copper
complex or a cobalt complex of hydroxycarboxylic acid may be led to
a chromophiric chelate compound respectively.
[0104] As a reagent which leads the inorganic salt of the
.beta.-hydroxycarboxylic acid compound to a chromophiric ester,
there may be p-nitro benzyl bromide, phenacyl bromide,
p-chlorophenacyl bromide, p-bromophenacyl bromide (PBPB),
p-iodophenacyl bromide, p-nitrophenacyl bromide, p-phenylphenacyl
bromide, p-phenylazophenacyl bromide,
N,N-dimethyl-p-aminobenzeneazophenacyl chloride or the like. As a
reagent which leads the hydroxamic acid to a chromophiric complex
salt, there may be ferric chloride, vanadium or the like. As a
reagent which leads the acid chloride to a chromophiric amide,
there may be 9-aminophenanthrene or the like. As a reagent which
leads the copper complex or cobalt complex of a hydroxycarboxylic
acid compound to a chromophiric chelate compound, there may be
diethyldithiocarbamate, bis(cyclohexanone)oxalyldihydrazone,
vasocupreine or the like. They may be used by optional choice as
required.
[0105] As a reagent which can be used in the gas chromatography
analysis, and reacts with a carboxyl group and/or a hydroxyl group
of the .beta.-hydroxycarboxylic acid compound, there may be a
silylation reagent such as N-trimethylsilylimidazole (TMSI),
N,O-bis (trimethylsilyl)acetamide (BSA) or the like, an acylation
reagent such as trifluoroacetic anhydride, trifluoroacetylimidazole
or the like.
[0106] The component(C) of the present invention may be subject to
chemical modification so as to be used as a substance (D) in order
to be detected in high sensitivity in the gas chromatography or
liquid chromatography or to be assessed by a spectrophotometer or
naked eye utilizing the coloring reaction unless it does not lose a
detection function as an indicator material. For example, an
atom(s) or atomic group(S) may be introduced to one or both of a
mercapto group and/or a hydroxyl group atom at the 3-position of a
3-mercapto alcohol compound and then used as derivatives such as
salt, ether, ester or the like.
[0107] As a reagent and method which and can be used in the liquid
chromatography analysis, spectrophotometer analysis and colorimetry
test and can be used for reacting with the mercapto group of a
3-mercapto alcohol compound, there may be a fluorescence method
which uses N-(9-acridinyl)maleimide (NAM),
4-chloro-7-sulfobenzofurazane ammonium salt (SBDCl),
4-fluoro-7-sulfobenzofurazane ammonium salt (SBD-F),
4-fluoro-7-sulfamoylbenzofrazane (ABD-F),
N-[4-(5,6-methylenedioxy-2-benzofuranyl)phenyl]maleimide (MBPM),
N-[4-(6-dimethylamino-2-benzofuranyl)phenyl]maleimide (DBPM),
N-[p-(2-benzimidazolyl)phenyl]maleimide, monobromobiman,
5,5-dithiobis (2-nitrobenzoic acid), phenanzine methosulfate,
o-phthal aldehyde, together with 2-aminoethanol, which may be used
by optional choice as required.
[0108] As a reagent which can be used in the liquid chromatography
analysis, spectrophotometer analysis or colorimetry test and can be
used for reacting with a hydroxyl group of a 3-mercapto alcohol
compound, there may be
3-chlorocarbonyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone
(DMEQ-COCl),
2-(5-chlorocarbonyl-2-oxazoyl)-5,6-methylenedioxybenzofuran,
3,4-dihydro-6,7-dimethoxy-4-methyl-3-oxoquinoxaline-2-carbony 1
chloride, phthalimidylbenzoyl chloride, 1-anthroylnitrile,
9-anthroylnitrile, 7-methoxycoumarin-3-carbonylazide,
p-phenylazobenzoyl chloride, 4-dialkylamino-3,5-dinitrobenzoyl
chloride, p-nitrobenzoyl chloride, 3,5-dinitrobenzoyl chloride or
the like.
[0109] As a reagent which can be used in the gas chromatography
analysis and can be used for reacting with a mercapto group and/or
hydroxyl group of a 3-mercapto alcohol compound, there may be a
silylation reagent such as hexamethyldisilazane (HMDS),
N-trimethylsilil imidazole or the like, an acylation reagent such
as trifluoroacetic anhydride, trifluoroacetylimidazole or the
like.
[0110] In the case of using a compound in which having a
chromophore of a visible region is introduced as a labeled compound
of the .beta.-hydroxycarboxylic acid compound and/or the 3-mercapto
alcohol compound, the level of apocrine odor can be assessed
visually by comparing the concentration-color standard sample which
has been preliminary prepared in relation to the concentration of
the labeled compound with coloration generated from perspiration of
a human using the same reagent.
[0111] The .beta.-hydroxycarboxylic acid compound and the
derivative thereof are suitable for the indicator material in terms
of capability in objective evaluation and assessment of the
apocrine odor at any time and anywhere since they can be
synthesized and the synthesized product with a certain quality can
be stably supplied. The .beta.-hydroxycarboxylic acid compound can
be synthesized, for example, according to the following formula
(5), wherein ester having a hydroxyl group at the .beta.-position
is synthesized by the Reformatsky Reaction disclosed in Reformatsky
Reaction: Ber. 20, 1210 (1887) and J. Russ. Phys. Chem. Soc., 22,
44 (1890), and the ester is hydrolyzed. ##STR9## wherein R.sup.1
and R.sup.2 are the same as aforementioned, and "X" is a halogen
atom.
[0112] Also, the 3-mercapto alcohol compound and the derivative
thereof are suitable for the indicator material in terms of
capability in objective evaluation and assessment of the apocrine
odor at any time and anywhere since they can be synthesized and the
synthesized product with a certain quality can be stably
supplied.
[0113] The 3-mercapto alcohol compound can be synthesized, for
example, according to the formula (6). That is, the 3-mercapto
alcohol compound is synthesized by preparing a derivative (a) of
fatty ester having unsaturated structure at the .beta.-position,
introducing benzylmercaptan or the like as a thio ether group to
the 3-position of the carbonyl carbon of the derivative (a) to
obtain a derivative (b) by the addition reaction, reducing the
derivative (b) with the use of a reducing agent such as lithium
aluminum hydride or the like to convert the ester group to an
alcohol group to obtain a derivative (c), and continuously leading
the thio ether group to a mercapto group by the Birch reduction.
##STR10## wherein R.sup.3, R.sup.4 and R.sup.5 are the same as the
formula (2), R.sup.6is a benzyl group and R.sup.7 is an alkyl
group.
[0114] The synthesized .beta.-hydroxycarboxylic acid compound and
3-mercapto alcohol compound may be converted as required to the
salt, ester or the other derivative by known means.
[0115] The .beta.-hydroxycarboxylic acid compound and 3-mercapto
alcohol compound, which have an asymmetric carbon atom, may be
synthesized as a racemic mixture, or each enantiomer may be
separately formed by asymmetric synthesis. Also, the racemic
mixture may be subjected to the optical resolution and thereafter
it may be used.
[0116] In the indicator material for assessing body odor of the
present invention, certainly the substance (A) and/or the substance
(C) itself can be used as an indicator component, derivatives of
the above mentioned indicator substances such as the substance (B)
and/or the substance (D) also may me used. Also, not derived
substance and a derived substance may be used in a combination. For
example, combinations of: the substances (A) and (D); the
substances (C) and (B); and the substances (A), (C) and (D) may be
used as an indicator material. Also, one or more kinds from each
group of substances (A) to (D) may be used in combination. Further,
one or more kinds may be selected from known causative agents of
the body odor of apocrine odor, acid odor or the like such as
acetic acid, butyric acid, isovaleric acid, 3-methyl-2-hexenoic
acid, 4-ethylheptanoic acid, 7-octenoic acid, 1-octene-3-one,
cis-1,5-octadiene-3-one, 3.alpha.-androstenol, 3.alpha.-androstenon
to be added to the indicator material for assessing body odor of
the present invention.
[0117] According to the indicator material for assessing body odor
of the present invention, it is possible to prepare an indicator
material which corresponds to various odors of the apocrine odor
different in feeling between cumin-like and sulfur-like by changing
weight ratio of the substance (A) and the substance (C) if
necessary. Particularly, from the view point of having the presence
ratio similar to the perspiration in the actual axillary regions,
it is preferable that the weight ratio in terms of "the substance
(C): the substance (A)" is 1:10 to 1:1,000 (weight ratio), more
preferably 1:10 to 1:500, and most preferably 1:50 to 1:200.
[0118] In the present invention, a body odor can be assessed with
the use of an indicator material of the present invention.
[0119] That is, in order to access the level of body odor of a
human or axillary odor which is a part of the body odor objectively
and quantitatively in terms of presence or strength of apocrine
odor of the axillary regions, the synthesized
.beta.-hydroxycarboxylic acid compound and/or the derivative
thereof can be used to determine quantity and observe the content
of the .beta.-hydroxycarboxylic acid compound and/or the derivative
thereof contained in the perspiration of the axillary regions.
Also, the synthesized 3-mercapto alcohol compound and/or the
derivative thereof can be used to determine quantity and observe
the content of the 3-mercapto alcohol compound and/or the
derivative thereof contained in the perspiration of the axillary
regions. Further, the synthesized .beta.-hydroxycarboxylic acid
compound and/or the derivative thereof and the synthesized
3-mercapto alcohol compound and/or the derivative thereof can be
used to determine quantity and observe the content of the
.beta.-hydroxycarboxylic acid compound and/or the derivative
thereof and the 3-mercapto alcohol compound and/or the derivative
thereof contained in the perspiration of the axillary regions.
[0120] Each of the selected indicator material may be solely used
as an indicator material or may be used as an indicator material
containing plural substance in mixture in the process of assessment
steps.
[0121] Also, in the present invention, body odor can be assessed by
using one or more kinds of .beta.-hydroxycarboxylic acid compound
and the derivative thereof and/or one or more kinds of 3-mercapto
alcohol compound and the derivative thereof as an index. That is,
"using as an index" in the present invention means to measure and
evaluate the substances (A), (B), (C) and/or (D) contained in the
perspiration. Typically, it means the case of using an indicator
material preliminarily prepared by synthesis or the like, however,
it may be a method which does not use an indicator material. For
example, it may be a method to measure the substances (A), (B), (C)
and/or (D) contained in the perspiration in a certain method and
evaluate the level of body odor or effectiveness of a deodorant
based on a standard data such as a calibration curve or the like
obtained preliminarily.
[0122] In order to measure the presence amount and the presence
state of the .beta.-hydroxycarboxylic acid compound and/or the
3-mercapto alcohol compound or the derivative thereof of the
present invention, there may be adapted some methods to obtain
perspiration originated from a human, such as a method that test
subjects is made to enter a room under the high temperature
environment and subjected to thermal sweating and then perspiration
of the axillary regions is collected in a test tube or the like, a
method that a cotton pad is attached to the axillary regions for a
certain time, a method that perspiration in the axillary regions is
wiped off by a cotton wool or the like.
[0123] The 3-mercapto alcohol compound is produced 10 to 100 times
or more by incubating the perspiration originated from a human
having the apocrine odor under the anaerobic atmosphere. On the
other hand, it is not produced even by incubating the perspiration
originated from a human who does not have an apocrine odor.
[0124] Utilizing this feature, analysis after incubation of the
obtained perspiration makes it easier to detect the 3-mercapto
alcohol compound, usefulness of the substance (C) as indicator
material enhances, and accuracy of the assessment result can be
enhanced at the same time. Further, by utilizing this feature, the
3-mercapto alcohol compound may be industrially produced as an
indicator material.
[0125] Also, if an apocrine sweat is secreting in the axillary
regions but the perspiration is not decomposed by microorganism and
the odor is not exhibited, it means that there is a latent state of
the apocrine odor. In such a state, an accurate evaluation cannot
be performed even if an organoleptic test or a research on the
relationship with an axillary odor is conducted.
[0126] On the contrary, by assessing body odor using an indicator
material of the present invention after incubating perspiration
originated from a human under an anaerobic atmosphere, it is
possible to evaluate if a test subject has a constitution which may
produce an apocrine odor, that is, it is possible to conduct a
potential evaluation.
[0127] As a method to prepare an anaerobic or microaerophilic
atmosphere for producing the 3-mercapto alcohol compound in large
quantity from perspiration collected, it is not particularly
limited as far as the method can remove oxygen in the incubation
atmosphere and replace with carbon dioxide. There may be methods
such as filling with mixed gas (nitrogen and carbon dioxide)
artificially prepared, using an agent which generates carbon
dioxide by vacuuming up oxygen gas or the like. Also, instead of
the binary mixed gas (nitrogen and carbon dioxide), a triple mixed
gas (nitrogen, carbon dioxide and hydrogen) may be used. As a
method of reducing concentration of remaining oxygen, there may be
a method wherein the remaining oxygen is absorbed by a reduction
steel wool, and a method which converts the oxygen into water using
catalyst.
[0128] The concentration of oxygen gas under an anaerobic or
microaerophilic atmosphere may be in the range of 0 to 10 v/v %
(volume in volume percent, hereinafter may be simply referred as
"%"), preferably 0 to 5%, more preferably 0 to 1%. Also, the
concentration of carbon dioxide may be in the range of 5.0 to
22.0%, preferably 10.0 to 22.0%, more preferably 20.0 to 22.0%.
[0129] The incubation temperature of perspiration originated from a
human for producing the 3-mercapto alcohol compound in large
quantity may be in the range of 15 to 45.degree. C., preferably 20
to 40.degree. C., more preferably 25 to 38.degree. C. Also, the
incubation time of perspiration may be in the range of 6 to 336
hours, preferably 12 to 240 hours, more preferably 24 to 168
hours.
[0130] As a method for assessing body odor of the present invention
using an indicator material for assessing body odor of the present
invention or using one or more kinds among the substances (A), (B),
(C) and (D) as an index, there may be a direct organoleptic test by
human olfaction and a quantitative evaluation based on the chemical
analysis, which may be used by adapting to various known evaluation
systems.
[0131] In the case of conducting an organoleptic test by olfaction,
an indicator material of the present invention in which
3-hydroxyl-3-methyl hexanoic acid or the derivative thereof and/or
the 3-mercapto alcohol compound or the derivative thereof is
diluted in several degrees to prepare odor standard samples of each
concentration. Then, odor of test sample prepared using the
perspiration collected from axillary regions and the standard
samples are matched to assess the amount of 3-hydroxyl-3-methyl
hexanoic acid and/or the 3-mercapto alcohol compound contained in
the perspiration by the organoleptic test.
[0132] In the case of measuring content of the substance (A) and
the substance (C) contained in the perspiration of the axillary
regions by GC-MS, 3-hydroxyl-3-methyl hexanoic acid or the
derivative thereof and the 3-mercapto alcohol compound or the
derivative thereof are preferably used as a standard material
(standard) to form each calibration curve. Using the calibration
curve, a peak of 3-hydroxyl-3-methyl hexanoic and the 3-mercapto
alcohol compound is respectively identified, and the amount is
measured.
[0133] In the case of using such an instrumental analysis, an
indicator material containing the substance (B) and the substance
(D) which are respectively derivatives obtained by reacting the
substance (A) and the substance (C) with the labeled material,
fluorescent reagent or the like which is easily detectable may be
used. Also, As a method for detecting the substance (C) and/or the
substance (D) which is the derivative thereof, there is a method to
introduce an extract of the obtained perspiration of a human as a
specimen using an organic solvent or the like directly to the gas
chromatography furnished with the highly sensitive sulfur
detector.
[0134] Also, as a measuring method for assessing body odor, a
coloration reagent may be added to the substance (A) and/or the
substance (C) separated from the perspiration so as to measure the
exhibited color by the spectrometer or conduct a colorimetry
assessment with the naked eye.
[0135] In the case of utilizing the coloration reaction, a visual
organoleptic test can be performed by preliminarily preparing
coloration standard samples of various concentration in a form of
colometric tubes filled with an aqueous solution or a form of test
papers impregnating labeled compound with the use of an indicator
material containing the substance (A) and the substance (C) reacted
with a coloration reagent or the like, and comparing the changed
color obtained by reacting the collected perspiration with the
coloration reagent with the coloration standard samples.
[0136] In this way, the level of body odor or axillary odor, which
is a part of the body odor, can be assessed by quantitatively
evaluating strength of the odor due to the substance (A) and
strength of the odor due to the substance (C) with the use of odor
or parameter other than odor and then assessing
comprehensively.
[0137] Also, if there is a production amount of 3-hydroxyl-3-methyl
hexanoic acid and/or the 3-mercapto alcohol compound is in large
quantity in the axillary regions but they are changed to a
derivative thereof which has no odor or weak odor such as a salt or
the like, it means that there is a latent state of the apocrine
odor. In such a state, an accurate evaluation cannot be performed
even if an organoleptic test or a research on the relationship with
an axillary odor is conducted. On the contrary, since derivatives
having latent odor of 3-hydroxyl-3-methyl hexanoic acid and/or and
the 3-mercapto alcohol compound can be analyzed and converted to
the quantity of the substance (A) and the substance (C) according
to the present invention, it is possible to evaluate if a test
subject has a constitution which may produce axillary odor, that
is, it is possible to conduct a potential evaluation.
[0138] An indicator material of the present invention can be
utilized for the chemical analysis, the instrumental analysis, the
organoleptic test or the like as described above and is capable of
quantitative assessment with high objectivity. Particularly, by
expressing the measured value as the quantity of the
.beta.-hydroxycarboxylic acid compound and/or the derivative
thereof and the 3-mercapto alcohol compound and/or the derivative
thereof with the use of the chemical analysis, instrumental
analysis or the like, the subjectivity can be excluded from the
assessment result and body odor can be assessed objectively and
quantitatively.
[0139] Further in the present invention, the effectiveness of a
deodorant targeting the apocrine odor can be assessed objectively
and quantitatively with the use of an indicator material of the
present invention or the substances constituting such a indicator
material as the index.
[0140] As a method for assessing effectiveness of a deodorant, the
indicator material may be used as a simple substance but also may
be used as a composition prepared by compounding other substances,
for example, a solvent for dissolution or dilution, or an additive
such as stabilizer, deodorant, bactericide, antibacterial agent,
surfactant, antioxidant, perfume, plant extracts or the like so as
to be adapted to the practical usage such as preservation, use in
assessment test or the like.
[0141] The deodorant targeting the apocrine odor may be of any
kinds of function mechanism such as a kind which prevents
decomposition of perspiration by disinfecting bacteria on the skin,
a kind which decomposes or changes an odor substance to an odorless
derivative, a kind which masks out odor or the like. A method for
using the .beta.-hydroxycarboxylic acid compound and/or the
derivative thereof, the 3-mercapto alcohol compound and/or the
derivative thereof, or both of them in combination as an indicator
material for assessing effectiveness of a deodorant may not be
particularly limited and may be used according to the function
mechanism of a deodorant and the evaluation system.
[0142] For example, the effectiveness of a deodorant sample can be
assessed objectively and quantitatively by adding a predetermined
amount of the deodorant sample to an indicator material containing
the .beta.-hydroxycarboxylic acid compound and/or the derivative
thereof, preferably 3-hydroxy-3-methylhexanoic acid or the
derivative thereof, and the 3-mercapto alcohol compound or the
derivative thereof, preferably 3-mercapto-3-methylhexanol,
3-mercaptohexanol, 3-mercaptopentanol, 3-mercapto-2-methylpentanol,
3-mercapto-2-methylbutanol or the derivatives thereof in a
predetermined concentration as the indicator substance, and
quantitatively determining the changed state of the indicator
material in an appropriate method.
[0143] As a method for quantitatively determining the changed state
of the indicator material, if the deodorant sample is a kind which
decomposes or leads the .beta.-hydroxycarboxylic acid and/or the
3-mercapto alcohol compound to other compound to reduce odor, an
instrumental analysis may be performed using a calibration curve of
the indicator material which is preliminarily formed or a chemical
analysis may be performed such as the titration or extraction of a
changed product or an unchanged state of the indicator material to
determine quantity. If the deodorant sample is a kind which masks
out the apocrine odor, the masking effect may be assessed by the
organoleptic test wherein the indicator material is diluted to
several levels to prepare odor standard samples of each
concentration and the odor of indicator material which is added
with the deodorant sample is matched to the standard samples.
[0144] Also, the quantitatively measurement may be conducted using
a labeled compound of the .beta.-hydroxycarboxylic acid and/or the
3-mercapto alcohol compound such as a fluorescent labeled compound,
in such manner that a predetermined amount of the deodorant sample
is added to an indicator material containing such a labeled
compound in a predetermined concentration and then the instrumental
analysis is performed for determining the changed state of the
indicator material with the use of the calibration curve of the
same indicator material. Also, a labeled portion of the labeled
compound to detect when a predetermined amount of deodorant sample
is added to an indicator material containing a labeled compound in
a predetermined concentration and thereafter the quantity of a
changed product or an unchanged state of the indicator material is
determined by the chemical analysis such as the titration,
extraction or the like.
[0145] Further, it is also possible to apply a deodorant sample
actually on the axillary regions of a human to evaluate and compare
each perspiration of the axillary regions obtained before and after
the application with the use of an indicator material of the
present invention.
[0146] In this way, effectiveness of a deodorant can be objevtively
and quantitatively assessed by quantitatively evaluating the
strength of odor due to the substance (A) and the strength of odor
due to the substance (C) contained in the apocrine odor of an
indicator material of the present invention in which a deodorant is
effected with the use of odor or parameter other than odor and then
assessing comprehensively.
[0147] As aforementioned, the level of body odor of a human or
axillary odor, which is a part of the body odor, can be objectively
and quantitatively assessed from the viewpoint of presence,
strength and difference in quality of the apocrine odor in the
axillary regions by using an indicator material for assessing body
odor containing at least one member selected from the group
consisting of the .beta.-hydroxycarboxylic acid compound
represented by the formula (1) (substance (A)), the derivative of
the substance (A) (substance (B)), the alcohol compound having a
mercapto group at the 3-position represented by the formula (2)
(substance (C)) and the derivative of the substance (C) (substance
(D)), or using at least one member selected from the group
consisting of the above-mentioned substances (A), (B), (C) and (D)
as an index.
[0148] Particularly, the odor due to the substance (A) and the odor
due to the substance (C) can be comprehensively evaluated by using
an indicator material for assessing body odor containing at least
one member selected from the group consisting of the substance (A)
and/or the substance (B) and at least one member selected from the
group consisting of the substance (C) and/or the substance (D), or
by using the substance (A) and/or (B) and the substance (C) and/or
(D) in combination as an index. Therefore, the apocrine odor which
humans actually smell can be more accurately assessed.
[0149] An indicator material for assessing body odor of the present
invention can objectively and quantitatively assess the body odor
particularly by expressing measured value as content of the
substance (A) and/or (B) or the substance (C) and/or (D) through
the chemical analysis, instrumental analysis or the like.
[0150] Even if the perspiration in the axillary regions is changed
to derivatives having no odor or weak odor such as the salt, ester
or the like of .beta.-hydroxycarboxylic acid compound and/or the
salt, ester or the like of 3-mercapto alcohol compound, the
.beta.-hydroxycarboxylic acid compound and/or the 3-mercapto
alcohol compound in a sample can be detected or quantitated by an
indicator material of the present invention. Therefore, the latent
state of axillary odor, namely, the state without odor or with weak
odor can be also objectively and accurately evaluated.
[0151] Also, if an apocrine sweat is secreting in the axillary
regions but the perspiration is not decomposed by microorganism and
the odor is not generated, it means that there is a latent state of
the apocrine odor. Even in such case, it is possible to evaluate if
a test subject has a constitution which may produce an apocrine
odor, that is, it is possible to conduct a potential evaluation by
incubating perspiration originated from a human according to the
present invention.
[0152] Further, in the present invention, an assessment result can
be accurately obtained corresponding to deodorizing and masking
effect against apocrine odor of a deodorant using an indicator
material containing a substance selected from the substances (A)
and (B) and/or a substance selected from the substances (C) and
(D), or using a substance selected from the substances (A) and (B)
and/or a substance selected from the substances (C) and (D) as an
index. Therefore, the effectiveness of a deodorant targeting
apocrine odor can be objectively and quantitatively assessed.
[0153] In the present invention, it is also possible to produce an
indicator material of the present invention by utilizing the
characteristic that the content of 3-mercapto alcohol compound
increases by the incubation of perspiration in the axillary regions
of a person having apocrine odor.
[0154] A kit for assessing body odor of the present invention is a
product which is a combination of at least a coloration reagent
which reacts with .beta.-hydroxycarboxylic acid originated from
perspiration of a human and accessories assisting a reaction of the
.beta.-hydroxycarboxylic acid and/or other substances causing body
odor originated from perspiration with the coloration reagent and
an assessment based on the coloration. In the accessories for
assistance, there are essential accessories such as extraction
means used for pretreatment which is always performed prior to the
coloration reaction or assessment, and accessories which improves
convenience such as facilitation, simplification or the like of the
coloration reaction or assessment.
[0155] A kit for assessing body odor includes, for example,
equipments and reagents for extracting or separating the
.beta.-hydroxycarboxylic acid and/or fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid from the
perspiration in the axillary regions, and one or more reagents
which exhibits-color by reacting with the .beta.-hydroxycarboxylic
acid and/or fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid originated from perspiration of a
human. Further, the kit may include 3-hydroxy-3-methylhexanoic acid
as an indicator substance.
[0156] FIG. 8 shows a schematic diagram of a process of assessing
body odor originated from perspiration of a human, particularly
apocrine odor, utilizing the coloration reaction of the
.beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid. Also, FIGS.
9 and 10 show typical equipment included in the kit for assessing
body odor and examples of the utility thereof.
[0157] As a method of collecting perspiration of the axillary
regions, there may be the method which directly collects
perspiration generated by the thermal sweating in test tubes or the
like, the method wherein test subjects wear T-shirt with cotton
pads sawn at a position corresponding to underarm for a certain
time, or the method which directly wipes perspiration in the
underarm with gauze or the like.
[0158] As a method of extracting the acid material contained in the
perspiration, it is not limited as far as the method can extract
the acid material. Generally, the acid-base extraction by alkali
aqueous solution is used. As the alkaline aqueous solution, there
may be sodium hydrogencarbonate aqueous solution, sodium carbonate
aqueous solution, sodium hydroxide aqueous solution, potassium
hydroxide aqueous solution or the like. It is also possible to
extract the acid material by using the ion-exchange resin.
[0159] As a method to assess a level of the apocrine odor, there
may be the method wherein the acid material extracted in the above
process is separated into .beta.-hydroxycarboxylic acid and fatty
acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid respectively, and thereafter a
coloration reagent is added to the separated
.beta.-hydroxycarboxylic acid to observe an exhibited color.
[0160] A method to separate .beta.-hydroxy carboxylic acid and the
fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid in the acid material is not
particularly limited as far as the method can separate using
polarity of molecule. There may be utilized the absorption
chromatography wherein a glass tube or plastic tube is filled with
an appropriate adsorbent such as silica gel powder or the like, the
thin-layer chromatography wherein a glass or plastic plate or the
like is coated with an appropriate adsorbent such as silica gel or
the like.
[0161] The absorption column chromatography can discharge in
ascending order of adsorbability to take up separately as
components having high adsorbability stays in the upper part and
components having lower adsorbability run off faster when the
solution of the above acid material is flowed off from the top of
the glass tube. In the case of using the acid material originated
from perspiration of a human as a sample, for example, as shown in
FIG. 9, after eluting fatty acid having 12 or less carbons other
than said .beta.-hydroxycarboxylic acid with the use of a middle
polar solvent such as diethyl ether, .beta.-hydroxycarboxylic acid
can be taken up with the use of a high polar solvent such as
methanol. Hence, when a coloration reagent is added to a methanol
fraction and the methanol fraction to generate coloration, the
presence of the .beta.-hydroxycarboxylic acid can be confirmed.
Also, the hue of test solution can be quantified by measurement
using equipments such as a spectrophotometer or color-difference
meter as well as the organoleptic test by the naked eye.
[0162] The thin-layer chromatography is the method which separates
molecules different in polarity on a plate, for example as shown in
FIG. 10, if the acid material originated from the perspiration of a
human is dropped in the predetermined position of a plate and then
developed with the use of the middle polar solvent such as diethyl
ether, the fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid spreads around together with the
solvent, however, .beta.-hydroxycarboxylic acid stays in the
dropped position. Hence, if a coloration reagent is dropped in the
center of the dropped spot to generate coloration, the presence of
.beta.-hydroxycarboxylic acid originated from perspiration can be
confirmed. In this manner, the thin-layer chromatography method can
directly spray the coloration reagent on the plate, therefore, the
qualitative or quantitative coloration test can be performed
quickly.
[0163] As a method which assess presence or quantity of
.beta.-hydroxycarboxylic acid using the coloration reaction, there
may be: (1) a method which directly introduces a chromophore to a
hydroxyl group or a carboxyl group of .beta.-hydroxycarboxylic
acid; (2) a method which converts .beta.-hydroxycarboxylic acid to
the derivative thereof and introduces a chromophore to the
derivative; (3) a method which decomposes .beta.-hydroxycarboxylic
acid and introduces a chromophore to the decomposed product; or the
like.
[0164] (1) As a coloration reagent which is used in a method which
directly introduces a chromophore to a hydroxyl group or a carboxyl
group of .beta.-hydroxycarboxylic acid and reacts with a carboxyl
group of .beta.-hydroxycarboxylic acid to exhibit color, there may
be a reagent which leads the .beta.-hydroxycarboxylic acid to a
chromophiric acid hydrazide in the presence of a condensing agent
to generate coloration, a reagent which leads the
.beta.-hydroxycarboxylic acid to a chromophiric ester to generate
coloration, a reagent which leads the .beta.-hydroxycarboxylic acid
to a chromophiric amide to generate coloration or the like.
[0165] As a coloration reagent which leads .beta.-hydroxycarboxylic
acid to the chromophiric acid hydrazide, there may be
2-nitrophenylhydrazine,
6,7-dimethoxy-1-methyl-2(1H)-quinoxaline-3-propionyl carboxylic
acid hydrazide (DMEQ-H),
p-(4,5-diphenyl-1H-imidazole-2-yl)-benzohydrazide,
p-(1-methyl-1H-phenanthro-[9,10-d]imidazole-2-yl)-benzo hydrazide,
p- (5,6-dimethoxy-2-benzothiazoyl)-benzohydrazide or the like.
[0166] As a reagent which leads .beta.-hydroxycarboxylic acid to
the chromophiric ester to generate coloration, there may be
9-anthryldiazomethane, 1-naphthyldiazomethane,
1-(2-naphthyl)diazomethane, 1-pyrenyldiazomethane,
4-diazomethyl-7-methoxycoumarin, 4-bromomethyl-7-methoxycoumarin,
3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone,
9-bromomethylacridine, 4-buromomethyl-6,7-methylenedioxycoumarin,
N-(9-acridinyl)-bromoacetamide,
2-(2,3-naphthylimino)ethyltrifluoromethane sulfonate,
2-(phthalimino)ethyltrifluoromethane sulfonate,
N-chloromethylphthalic imide, N-chloromethyl-4-nitrophthalic imide,
N-chloromethylisatin, o-(p-nitrobenzyl)-N,N'-diisopropylisourea or
the like.
[0167] As a reagent which leads the 3-hydroxycarboxylic acid to the
chromophiric amide to generate coloration, there may be
monodansylcadaverine,
2-(p-aminomethylphenyl)-N,N-dimethyl-2H-benzotriazole-5-amine or
the like.
[0168] As a coloration reagent which reacts with a hydroxyl group
of .beta.-hydroxycarboxylic acid to generate coloration, there may
be a reagent which leads to a chromophiric coordination compound to
generate coloration, a reagent which leads to a chromophiric ester
to generate coloration, a reagent which leads to a chromophiric
ether to generate coloration or the like. As a reagent which leads
to the chromophiric coordination compound to generate coloration,
there may be cerium nitrate ammonium or the like. As a reagent
which leads to the chromophiric ester to generate coloration, there
maybe 4-(2-phthalimidyl)benzoyl chloride, the isomer there of, the
derivative there of or the like. As a reagent which leads to the
chromophiric ether to generate coloration, there may be
4-diazomethyl-7-methoxycoumarin or the like.
[0169] (2) As a derivative of .beta.-hydroxycarboxylic acid which
can be utilized for the coloration reaction in the method which
converts .beta.-hydroxycarboxylic acid to the derivative thereof
and then introduces a chromophore to the derivative, there may be
inorganic salt, hydroxamic acid, acid chloride, copper complex,
cobalt complex or the like.
[0170] The inorganic salt of .beta.-hydroxycarboxylic acid reacts
with aromatic halogen to be led to the chromophiric ester,
hydroxamic acid is led to the chromophiric metal salt, acid
chloride is led to the chromophiric amide, a copper complex and a
cobalt complex of the hydroxycarboxylic acid are led to a chelate
compound which react with copper and cobalt respectively to
generate coloration.
[0171] As a method to convert .beta.-hydroxycarboxylic acid to
inorganic salt, there may be a method which mixes the
.beta.-hydroxycarboxylic acid with the alkaline material such as
sodium hydrogen carbonate solution, sodium carbonate solution,
sodium hydroxide solution, potassium hydroxide solution or the like
to neutralize. As an aromatic halogen which reacts with inorganic
salt of the hydroxycarboxylic acid to be led to the chromophiric
ester, there may be p-nitrobenzyl bromide, phenacyl bromide,
p-chlorophenacyl bromide, p-bromophenacyl bromide, p-iodinephenacyl
bromide, p-nitrophenacyl bromide, p-phenylphenacyl bromide,
p-phenylazophenacyl bromide, N,N-dimethyl-p-aminobenzeneazophenacyl
chloride or the like.
[0172] As a method to convert .beta.-hydroxy carboxylic acid to
hydroxamic acid, there may be a method which reacts .beta.-hydroxy
carboxylic acid with hydroxylamine in the presence of a condensing
agent, a method which reacts .beta.-hydroxy carboxylic acid with
hydroxylamine hydrochloride using nickel as catalyst or the like.
As a metal reagent which forms the chromophiric complex salt with
hydroxamic acid, there may be ferric chloride, vanadium or the
like.
[0173] As a method to convert .beta.-hydroxycarboxylic acid to acid
chloride, there may be a method which reacts
.beta.-hydroxycarboxylic acid with oxalylchloride or the like. As a
method to lead acid chloride to the chromophiric amide, there may
be a method which reacts it with 9-aminophenanthrene in the
presence of triethylamine or the like.
[0174] As a reagent which forms the chromophiric chelate compound
(complex) with copper or cobalt, there may be diethyldithiocarbamic
acid, bicyclohexanoneoxalyldihydrazone, bathocuproin or the
like.
[0175] (3) As a method which decomposes .beta.-hydroxycarboxylic
acid and makes the decomposed product exhibit color, there may be a
method wherein acyl-CoA synthetase is effected to
.beta.-hydroxycarboxylic acid in the presence of adenosine
triphosphoric acid (ATP) and coenzyme CoA to produce acyl-CoA,
which is then processed with acyl-CoA oxidase to produce enoyl-CoA
and hydrogen peroxide, the hydrogen peroxide is further processed
with catalase to obtain formaldehyde, and the obtained formaldehyde
is reacted with 4-amino-3-hydrazino-5-mercapto-1,2,3-triazole
(AHMT), which is a coloration reagent, to exhibit purple color
which is used for the colorimetry test.
[0176] In this manner, the reagent used for the coloration reaction
of .beta.-hydroxycarboxylic acid in the present invention may not
be particularly limited as far as the reagent reacts with
.beta.-hydroxycarboxylic acid, the derivative of
.beta.-hydroxycarboxylic acid or the decomposed product of
.beta.-hydroxycarboxylic acid. A compound having a hydrazino group
such as 2-nitrophenylhydrazine (2-NPH) or the like, which has high
sensitivity in detecting .beta.-hydroxycarboxylic acid originated
from the perspiration of a human and is easy to perform the
colorimetry test with the naked eye, or a compound having a
diazomethyl group such as 9-anthryldiazomethane (ADAM) or the like,
which has high sensitivity in detecting and reacts under a mild
condition, are suitably used.
[0177] The coloration reaction using 2-nitrophenylhydrazine is a
reaction (Formula (7)) in which .beta.-hydroxycarboxylic acid
reacts with 2-nitrophenylhydrazine in water or alcohol solution in
the presence of a condensing agent to produce acid hydrazide which
exhibits magenta under the alkali condition: ##STR11##
[0178] As a condensing agent, there may be utilized,
dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide (EDC) or the like. As an alkaline agent, there may be
utilized potassium hydroxide solution or the like.
[0179] When 0.5 mL of water or alcohol solution containing 0.02M
2-nitrophenylhydrazine hydrochloride is added to 0.5 mL of water or
alcohol solution containing 0.01 to 0.05 .mu.M
3-hydroxy-3-methylhexanoic acid, the test solution exhibits
slightly brownish yellow. Also, when 0.5 mL of water or alcohol
solution containing 0.02M 2-nitrophenylhydrazine is added to 0.5 mL
of water or alcohol solution containing 0.05 to 1 .mu.M
3-hydroxy-3-methylhexanoic acid, the test solution exhibits sienna
to dark magenta. On the contrary, when water or alcohol solution
containing 2-nitrophenylhydrazine is added to water or alcohol
solution not containing hydroxycarboxylic acid, acid hydrazide is
not produced and the test solution remains yellow, which is an
original color of unreacted 2-nitrophenylhydrazine.
[0180] As aforementioned, the strength of the apocrine odor is
proportional to the content of .beta.-hydroxycarboxylic acid in the
perspiration. Hence, a level of apocrine odor can be surely,
quickly and easily assessed by observing the developed color of a
test solution with the naked eye.
[0181] The coloration reaction using 9-anthryldiazomethane is a
method to lead .beta.-hydroxycarboxylic acid to 9-anthrylmethyl
ester having strong fluorescence (Formula (8)): ##STR12##
[0182] The reaction does not need a catalyst or heat and finishes
in about 10 to 60 minutes at room temperature. 9-Anthrylmethyl
ester in methanol has the excitation wavelength of 365 nm and the
fluorescent wavelength of 412 nm.
[0183] When 1.0% 9-anthryldiazomethane in methanol or acetone
solution is added to methanol solution containing 0.01 to 0.05
.mu.M 3-hydroxy-3-methylhexanoic acid followed by radiation with
light of 365 nm, the test solution exhibits slightly bluish white.
Further, when 1.0% 9-anthryldiazomethane in methanol or acetone
solution is added to methanol solution containing 0.05 to 1 .mu.M
3-hydroxy-3-methylhexanoic acid followed by radiation with light of
365 nm, the test solution exhibits strong blue fluorescence. On the
contrary, even though 1.0% 9-anthryldiazomethane in methanol
alcohol solution is added to a solution not containing
.beta.-hydroxycarboxylic acid, an ester having strong fluorescence
is not produced.
[0184] As aforementioned, the strength of the apocrine odor is
proportional to the content of .beta.-hydroxycarboxylic acid in the
perspiration. Hence, a level of apocrine odor can be surely,
quickly and easily assessed by observing the developed color of a
test solution with the naked eye.
[0185] In the present invention, when .beta.-hydroxycarboxylic acid
contained in the perspiration is separated from the perspiration of
the axillary regions and thereafter quantitative evaluation is
performed using a coloration reagent, synthesized
3-hydroxy-3-methylhexanoic acid can be used as a standard material
(standard).
[0186] That is, the amount of .beta.-hydroxycarboxylic acid
contained in the perspiration can be more accurately determined by
using the exhibited color of 3-hydroxy-3-methylhexanoic acid which
is synthesized and weighed and a coloration reagent as standard. At
the same time, the amount of .beta.-hydroxycarboxylic acid is
proportional to the strength of the apocrine odor, thus, a level of
the apocrine odor can be assessed accurately.
[0187] The standard material, 3-hydroxy-3-methylhexanoic acid, used
therein may be diluted to proper concentration in a laboratory,
however, it is convenient if the standard material is diluted in a
solvent such as methanol, acetone or the like so as to facilitate
to take up a suitable amount for comparison. Particularly, it is
preferable to use a standard solution diluted to the degree of
being able to measure off, such as a diluted solution of About 1 to
1,000 .mu.g 3-hydroxy-3-methylhexanoic acid in 100 .mu.L
diluent.
[0188] As aforementioned, the .beta.-hydroxycarboxylic acid
compound and the derivative thereof such as
3-hydroxy-3-methylhexanoic acid or the like can be synthesized by
utilizing the Reformatsky reaction.
[0189] Also, in the present invention, if the difference in color
between samples to compare is subtle or when assessing a level of
apocrine odor reduced after the use of a deodorant or an operation
to remove apocrine glands, it is possible to exclude objectivity
from the assessment result by quantifying hue of the test solution
with the use of an analytical instrument.
[0190] The analytical instrument used therein is not particularly
limited as far as the instrument can measure the degree of
exhibited color of the test solution. There may be used a
calorimeter, ultraviolet-visible spectrophotometer or the like for
acid hydrazide produced by the coloration reaction using
2-nitrophenyl hydrazine. Also, a fluorospectrophotometer or the
like may be utilized for 9-anthrylmethyl ester produced by the
coloration reaction using 9-anthryldiazomethane.
[0191] Further in the present invention, not only a level of
apocrine odor specifically generated to the axillary regions of a
person having tragomaschalia habit but also a level of acid odor
(perspiration odor) generated in the axillary regions of a person
not depending on having tragomaschalia habit can be quickly and
easily assessed.
[0192] As a method for assessing a level of contribution of
apocrine odor and acid odor, there may be a method wherein after
separating an acid material extracted from perspiration into
.beta.-hydroxycarboxylic acid, which causes apocrine odor, and
fatty acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid, a coloration reagent is added
thereto respectively to observe the exhibited colors, or a method
wherein a coloration reagent is added to the acid material
extracted from perspiration and .beta.-hydroxycarboxylic acid
separated from the acid material respectively to observe the
exhibited colors.
[0193] As a method for assessing a level of acid odor, a coloration
reagent may be added to fatty acid having 12 or less carbons other
than said .beta.-hydroxycarboxylic acid, which causes acid odor.
For example, as shown in FIG. 9, in the adsorption column
chromatography, a level of contribution of apocrine odor and acid
odor can be quickly and easily assessed by eluting acid component
originated from perspiration, which is mainly fatty acid having 12
or less carbons other than said .beta.-hydroxycarboxylic acid with
the use of diethyl ether as a developing solvent, thereafter
eluting .beta.-hydroxycarboxylic acid with the use of methanol, and
then adding a coloration reagent to the obtained ether fraction and
methanol fraction respectively to observe the exhibited color.
[0194] Also, in the thin-layer chromatography, the acid material
extracted from perspiration of a human is dropped on a plate for
the thin-layer chromatography to which a coloration reagent is
dropped to let a whole acid material exhibit color. The color
exhibited by a whole acid material has a positive correlation with
respect to the total amount of .beta.-hydroxycarboxylic acid, which
is a component causing the apocrine odor, and fatty acid having 12
or less carbons other than said .beta.-hydroxycarboxylic acid,
which is a component causing the acid odor.
[0195] On the other hand, after dropping the extracted acid
material on a similar plate for the thin-layer chromatography
prepared separately and separating .beta.-hydroxycarboxylic acid
and fatty acid other than said .beta.-hydroxycarboxylic acid with
the use of the developing solvent, the coloration reagent is
dropped on the .beta.-hydroxycarboxylic acid remained in the center
of the dropped spot to exhibit color. The color exhibited on this
plate has a positive correlation with respect to the amount of
.beta.-hydroxycarboxylic acid, which is a major component causing
the apocrine odor. Thus, by observing both exhibited colors, a
level of contribution of apocrine odor and acid odor can be
assessed.
[0196] Therefore, it is possible to assess a level of contribution
of apocrine odor and acid odor of each test subject quickly and
easily with the use of a kit for assessing body odor of the present
invention.
[0197] As a coloration reagent which reacts with carboxylic acid
contained in perspiration to generate coloration, which may not
particularly limited as far as the coloration reagent reacts with a
carboxyl group of fatty acid having 12 or less carbons, which has
major contribution to acid odor, there may be utilized the
aforementioned coloration reagent which reacts with a carboxyl
group of .beta.-hydroxycarboxylic acid to generate coloration.
[0198] In order to assess the kind and strength of body odor
accurately, i.e. for an accurate assessment of the degree of each
contribution of apocrine odor and acid odor with respect to the
acid material originated from perspiration, it is preferable that
the reactivity of coloration reagent is highly specific to
.beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid and the
coloration reaction with respect to other acid material is
weak.
[0199] As aforementioned, a kit for assessing body odor of the
present invention can surely, quickly and easily assess the level
of body odor or comprehensive axillary odor, which is a part of
body odor, focusing on presence and strength of apocrine odor of
the axillary regions and further taking the presence and strength
of acid odor into account without using expensive analytical
instruments such as the gas chromatography, liquid chromatography
or the like. Therefore, the kit can be utilized when conducting
diagnosis on a level of apocrine odor of a person who is concerned
about own axillary odor, or assessing effectiveness of a deodorant,
an operation to remove apocrine glands or the like at inspection
institutes such as hospital (dermatology or the like), health
center or the like.
[0200] In the case of assessing whether an effort to reduce body
odor such as the use of a deodorant or an operation to remove
apocrine glands is actually effective, for example, the quantity of
.beta.-hydroxycarboxylic acid and/or fatty acid having 12 or less
carbons other than said .beta.-hydroxycarboxylic acid in the
perspiration before and after the use of a deodorant may be
determined and evaluated using a kit for assessing body odor of the
present invention to compare. For example, if the time of
collecting each sample is different, the first collected
perspiration sample may be stored in an appropriate place such as a
refrigerator so that the sample does not cause chemical change and
the assessment test may be performed at the same time. When the
quantity of .beta.-hydroxycarboxylic acid and fatty acid having 12
or less carbons other than said .beta.-hydroxycarboxylic acid in
the perspiration is determined and evaluated, if the colors of test
solutions are quantified by an analytical instrument, comparison
and evaluation can be performed accurately even though the time
when the assessment test is performed is different.
[0201] Also, a kit for assessing body odor of the present invention
can be utilized for evaluating effectiveness of a deodorant sample
or screening people having tragomaschalia habit at development
research of deodorants targeting apocrine odor.
[0202] As a method for assessing effectiveness of a deodorant
targeting apocrine odor, for example, the effectiveness of a
deodorant sample can be assessed precisely, rapidly and easily by
adding predetermined amount of a deodorant sample to a sample
containing .beta.-hydroxycarboxylic acid of a predetermined
concentration, and assessing the amount of .beta.-hydroxycarboxylic
acid, preferably the amount of 3-hydroxy-3-methylhexanoic acid,
with the use of a kit for assessing body odor of the present
invention.
[0203] It is also possible to apply a deodorant sample actually on
the axillary regions of a human and evaluate
.beta.-hydroxycarboxylic acid, preferably the amount of
3-hydroxy-3-methylhexanoic acid, contained in perspiration of the
axillary regions collected respectively before and after the
application with the use of a kit for assessing body odor of the
present invention to compare.
[0204] As example of the constitution of the aforementioned kit,
for example, there may be the followings, but may not be limited
thereto.
[0205] (1) A package including a cotton wool, plastic cup or the
like for collecting perspiration of a human, preferably processed
to be usable without modification, and wrapped with an outer
package as required;
[0206] (2) A package including reagents, solvents and equipments
such as an aqueous solution of an alkali, organic solvent,
separating funnel or the like for extracting acid material from the
perspiration of a human in combination with the above
constituents;
[0207] (3) A package including equipments such as columns for the
adsorption chromatography, plates for the thin-layer chromatography
or the like to separate .beta.-hydroxycarboxylic acid and fatty
acid having 12 or less carbons other than said
.beta.-hydroxycarboxylic acid originated from perspiration of a
human in combination with constituents of any of the above
packages;
[0208] (4) A package including middle polar solvents used for
eluting fatty acid having 12 or less carbons other than the
.beta.-hydroxycarboxylic acid from acid material in the adsorption
chromatography or the thin-layer chromatography in combination with
constituents of any of the above packages;
[0209] (5) A package including high polar solvents used for eluting
.beta.-hydroxycarboxylic acid from a column in the adsorption
chromatography in combination with constituents of any of the above
packages;
[0210] (6) A package including a coloration reagent which reacts
with .beta.-hydroxycarboxylic acid originated from perspiration of
a human, preferably filled in a container in which the coloration
reagent is prepared in the condition of composition or
concentration which can be used without modification in combination
with constituents of any of the above packages and wrapped with an
outer package as required;
[0211] (7) A package including a coloration reagent which reacts
with fatty acid having 12 or less carbons other than the
.beta.-hydroxycarboxylic acid, preferably filled in a container in
which the coloration reagent is prepared in the condition of
composition or concentration which can be used without modification
in combination with constituents of any of the above packages and
wrapped with an outer package as required;
[0212] (8) A package including a coloration reagents which react
with .beta.-hydroxycarboxylic acid and fatty acid having 12 or less
carbons other than the .beta.-hydroxycarboxylic acid originated
from perspiration of a human, preferably filled in a container in
which the coloration reagents are prepared in the condition of
composition or concentration which can be used without modification
in combination with constituents of any of the above packages and
wrapped with an outer package as required;
[0213] (9) A package including auxiliary reagents used for
preliminary preparations prior to the coloration reaction when the
quantity of .beta.-hydroxycarboxylic acid is determined and
evaluated using a coloration reagent after the
.beta.-hydroxycarboxylic acid is separated from perspiration of the
axillary regions, for example, a preliminary preparation to lead
the .beta.-hydroxycarboxylic acid to the derivative, in combination
with constituents of any of the above packages;
[0214] (10) A package including equipments such as ultraviolet
lamps used for confirming a level of coloration of a test solution
when the quantity of .beta.-hydroxycarboxylic acid is determined
and evaluated using a coloration reagent after the
.beta.-hydroxycarboxylic acid is separated from perspiration of the
axillary regions in combination with constituents of any of the
above packages;
[0215] (11) A package including a diluted solution of
3-hydroxy-3-methylhexanoic acid or 3-hydroxy-3-methylhexanoic acid
which are preferably synthesized products and used as standard
material (standard) when the quantity of .beta.-hydroxycarboxylic
acid contained in perspiration is determined and evaluated using a
coloration reagent after said .beta.-hydroxycarboxylic acid is
separated from perspiration of the axillary regions in combination
with constituents of any of the above packages;
[0216] (12) A package including color sample sheets for assessing a
level of coloration of test solutions when the quantity of
.beta.-hydroxycarboxylic acid contained in perspiration is
determined and evaluated using a coloration reagent after the
.beta.-hydroxycarboxylic acid is separated from perspiration of the
axillary regions in combination with constituents of any of the
above packages;
[0217] (13) A package including means for dropping such as a
pipette or the like which is easy to take up and drop a single dose
of a coloration reagent in combination with constituents of any of
the above packages; and
[0218] (14) A package including analytical instrument such as the
calorimeter, spectrometer or the like used for quantifying a level
of exhibited color of test solutions in combination with
constituents of any of the above packages.
[0219] A kit for assessing body odor of the present invention can
surely, quickly and easily assess the level of body odor or
axillary odor, which is a part of body odor, focusing on presence
and strength of apocrine odor of the axillary regions and further
taking the presence and strength of acid odor into account,
utilizing the coloration reaction of .beta.-hydroxycarboxylic acid,
which is peculiarly present in perspiration of a person having
apocrine odor, and/or fatty acid having 12 or less carbons other
than the .beta.-hydroxycarboxylic acid, which is a material causing
acid odor or the derivative thereof or the decomposed product
thereof.
[0220] Also, a kit for assessing body odor and a method for
assessing body odor of the present invention can exclude
objectivity from the assessment result by quantifying hue exhibited
as the result of the coloration reaction of the test solution with
the use of an analytical instrument such as a calorimeter or the
like.
[0221] Further, a kit for assessing body odor and a method for
assessing body odor of the present invention can be utilized as a
sure, quick and easy kit or method for assessing body odor when
conducting diagnosis on a level of apocrine odor of a person who is
concerned about own axillary odor, assessing effectiveness of an
operation to remove apocrine glands or the like, assessing
effectiveness of a deodorant sample targeting apocrine odor or the
like at a hospital, health center, research institute or the
like.
EXAMPLE
[0222] The following examples further describe and demonstrate
embodiments of the present invention. The examples are given solely
for the purpose of illustration and are not to be construed as
limitations of the present invention.
Example A
1. Screening of Test Subjects
[0223] Each of 65 healthy Japanese males selected at random as test
subjects wore a new white T-shirt made of cotton for 24 hours
consecutively and the T-shirts were collected. An organoleptic test
by 7 professional panel members was conducted on the axillary
regions of the collected T-shirts concerning kinds and strength of
axillary odor.
<The Points of Evaluation for Odor>
[0224] Acid odor: sour and putrefactive odor
[0225] Cumin-like apocrine odor: spicy, woody and animal-like
odor
[0226] Sulfur-like apocrine odor: fishy and soy sauce and
grapefruit-like odor
<The Evaluation Criteria of for Strength of Odor>
[0227] 0: Odorless
[0228] 1: Slight odor
[0229] 2: Weak odor
[0230] 3: Easily perceptible odor
[0231] 4: Strong odor
[0232] 5: Very strong odor
[0233] As a result, 28 people were recognized to have little
axillary odor (the strength of odor of these people was 1 or less)
(herein after referred as Group A), 21 people were recognized to
have weak axillary odor (the highest strength of odor of these
people was 2 or 3) (herein after referred as Group B), and 16
people were recognized to have strong axillary odor (the highest
strength of odor of these people was 4 or 5) (herein after referred
as Group C).
[0234] The evaluation result of the Group C, which was recognized
to have strong axillary odor, is shown in Table 1. As shown in
Table 1, the test subjects having apocrine odor can be classified
into a group shifting to the cumin-like odor (Subject
identification Nos. 1 to 3) and a group shifting to the sulfur-like
odor (Subject identification Nos. 4 to 9). TABLE-US-00001 TABLE 1
Organoleptic evaluation result of Group C Organoleptic evaluation
result Subject Apocrine odor identification Lower fatty Cumin-like
Sulfur-like No. acid odor odor odor 1 3 5 4 2 3 5 3 3 2 5 4 4 4 4 5
5 4 4 5 6 3 3 5 7 3 3 4 8 2 3 4 9 3 3 4 10 4 2 2 11 4 2 1 12 4 2 1
13 4 1 2 14 5 0 0 15 4 0 0 16 4 0 0
2. Analysis of Component Corresponding to Cumin-Like Odor
[0235] The test subjects were 13 people who were recognized to have
a cumin-like apocrine odor in the Group C (Subject identification
Nos. 1 to 13). A cotton pad was sewn on a part of a T-shirt
corresponding to the axillary regions. After the test subjects wore
the T-shirts for 24 hours, the T-shirts were collected and the
cotton pad of the axillary region was subject to extraction in
diethyl ether. The extract was divided into an acid fraction, a
neutral fraction and a basic fraction by an acid-base extraction
method, and volatile components were analyzed with the used of a
Gas Chromatography-Mass Spectrometer (GC-MS). Important components
which generate typical odors of perspiration of an axillary region
were specified by a sniffing Gas Chromatography (sniffing GC).
<GC-MS Analysis Conditions>
[0236] Device: 6890GC-5973MSD (Agilent Technology)
[0237] Column: DB-WAX (60 m.times.0.25 mm.times.0.25 .mu.m)
[0238] Temperature conditions: 40.degree. C. (1
minute).fwdarw.(6.degree. C./min.).fwdarw.60.degree.
C..fwdarw.(2.degree. C./min.).fwdarw.300.degree. C. (40
minutes)
[0239] Career gas: He
[0240] Ionization voltage: 70 eV
[0241] By analyzing the acid extract, a new presence of
3-hydroxy-3-methylhexanoic acid was shown together with
conventionally known presence of saturated fatty acid,
3-methyl-2-hexenoic acid, 7-octenoic acid and .gamma.-lactones
(FIG. 1). This component had a strong odor very similar to the
apocrine odor at the time of performing the sniffing GC. Also, the
amount of 3-hydroxy-3-methylhexanoic acid contained in the acid
extract was calculated from a peak area and it was in the range of
0.1 to 64.3 .mu.g. It was found that 3-hydroxy-3-methylhexanoic
acid is present at high concentrations capable of being
quantitatively detectable. 3-hydroxy-3-methylhexanoic acid has not
reported as a constituent of an axillary odor of a human so far,
however, 3-hydroxy-3-methylhexanoic acid was detected from all the
test subjects.
[0242] Next, test subjects were 3 people who were recognized to
have no apocrine odor in the Group C (Subject identification Nos.
14 to 16). A cotton pad was sewn on a part of a T-shirt
corresponding to the axillary region. After the test subjects wore
the T-shirts for 24 hours, the T-shirts were collected and the
cotton pad of the axillary region was subject to extraction in
diethyl ether. An acid fraction was extracted by an acid-base
extraction method, and analyzed with the used of a Gas
Chromatography-Mass Spectrometer (GC-MS).
[0243] As a result, as shown in FIG. 2, 3-hydroxy-3-methylhexanoic
acid was not detected from any test subject.
3. Determining Quantity of 3-hydroxy-3-methylhexanoic Acid
[0244] Test subjects were 5 people who were recognized to have
strong apocrine odors in the Group C (Subject identification Nos. 1
to 5), 4 people who were recognized to have weak apocrine odors in
the Group C (Subject identification Nos. 10 to 13) and 3 people who
were recognized to have no apocrine odor in the Group C (Subject
identification Nos. 14 to 16). A cotton pad was sewn on a part of a
T-shirt corresponding to the axillary region. After the test
subjects wore the T-shirts for 24 hours, the T-shirts were
collected and the cotton pad of the axillary region was subject to
extraction in diethyl ether. An acid fraction was extracted by an
acid-base extraction method. Then, a diluted solution using ether
of the same volume was prepared by a 1 ml measuring flask
(manufactured by DRAN). Using the diluted solution, the quantity of
3-hydroxy-3-methylhexanoic acid was determined with the use of a
Gas Chromatography-Mass Spectrometer (GC-MS). A quantitative value
was calculated from a peak area which was obtained by injecting
3-hydroxy-3-methylhexanoic acid having a given concentration to the
GC-MS.
[0245] The relationship between the presence or level of apocrine
odor and the detected amount of 3-hydroxy-3-methylhexanoic acid was
obtained. As a result, as shown in FIG. 3, the amount of
3-hydroxy-3-methylhexanoic acid at the axillary region increased
when the apocrine odor became stronger.
4. Analysis of Component Corresponding to Sulfur-Like Odor
[0246] The test subjects were 3 people who were recognized to have
a sulfur-like apocrine odor in the Group C (Subject identification
Nos. 4 to 6). About 1 mL of perspiration running from both axillary
regions of each test subject was collected in a test tube in a room
conditioned at 40.degree. C. and 80% humidity. The test tube in
which Twister (a stirrer coated with 100% polydimethylsiloxane,
alias Stir Bar Sorptive Extraction; manufactured by Gerstel) was
added was agitated for 30 minutes. Then, an analysis was performed
with the use of a Gas Chromatography-Mass Spectrometer (GC-MS)
equipped with a heat desorption device. Important components which
generate typical odors of perspiration of an axillary region were
specified by a sniffing Gas Chromatography (sniffing GC) equipped
with a heat desorption device.
<GC-MS Analysis Conditions>
[0247] Device: 6890GC-5973MSD (Agilent Technology)
[0248] Column: DB-1 (60 m.times.0.25 mm.times.0.25 .mu.m)
[0249] Temperature conditions: 40.degree. C. (1
minute).fwdarw.(6.degree. C./min.).fwdarw.60.degree.
C..fwdarw.(2.degree. C./min.).fwdarw.300.degree. C. (40
minutes)
[0250] Career gas: He
[0251] Ionization voltage: 70 eV
[0252] By analyzing the components of the apocrine odor contained
in perspiration, a new presence of 3-mercapto-3-methylhexanol was
shown by GC-MS analysis (FIG. 4). The eluted component had a strong
odor very similar to the apocrine odor at the time of performing
the sniffing GS. Also, the amount of 3-mercapto-3-methylhexanol was
calculated from a peak area and it was in the range of 0.001 to 1.0
.mu.g. It was found that the 3-mercapto-3-methylhexanol is present
at high concentrations capable of being quantitatively
detectable.
[0253] 3-mercapto-3-methylhexanol has not reported as a constituent
of an axillary odor of a human so far, however,
3-mercapto-3-methylhexanol was detected from all 3 test
subjects.
[0254] Next, test subjects were 3 people who were recognized to
have no apocrine odor in the Group C (Subject identification Nos.
14 to 16). About 1 mL of perspiration running from both axillary
regions of each test subject was collected in a test tube in a room
conditioned at 40.degree. C. and 80% humidity. The test tube in
which the Twister was added was agitated for 10 minutes. Then, an
analysis was performed with the use of a Gas Chromatography-Mass
Spectrometer (GC-MS) equipped with a heat desorption device. As a
result, as shown in FIG. 5, 3-mercapto-3-methylhexanol was not
detected from any test subject.
[0255] Test subjects were 3 people who were recognized to have a
strong sulfur-like apocrine odor in the Group C (Subject
identification Nos. 4 to 6) and 3 people who were recognized to
have no apocrine odor in the Group C (Subject identification Nos.
14 to 16). About 1 mL of perspiration running from both axillary
regions of each test subject was collected in a test tube in a room
conditioned at 40.degree. C. and 80% humidity. After the
perspiration was incubated in an anaerobic environment (an oxygen
concentration of 0.1% or less and a carbon dioxide concentration of
21%) at 30.degree. C. for 48 hours, the test tube in which the
Twister was added was agitated for 10 minutes. Then, an analysis
was performed with the use of a Gas Chromatography-Mass
Spectrometer (GC-MS) equipped with a heat desorption device.
Important components which generate typical odors of perspiration
of an axillary region were specified with the use of a sniffing Gas
Chromatography (sniffing GC) equipped with a heat desorption
device.
[0256] As a result, as shown in FIG. 6, 3-mercapto-3-methylhexanol,
3-mercaptohexanol, 3-mercaptopentanol, 3-mercapto-2-methylpentanol
and 3-mercapto-2-methylbutanol were identified from the incubated
perspiration of people having an apocrine odor. Each of these
eluted components had a strong odor similar to the apocrine odor at
the time of performing the sniffing GC, which became stronger by
the incubation.
[0257] On the contrary, 3-mercapto-3-methylhexanol,
3-mercaptohexanol, 3-mercaptopentanol, 3-mercapto-2-methylpentanol
and 3-mercapto-2-methylbutanol were not detected from the
perspiration of any test subject having no apocrine odor even it
was incubated (FIG. 7).
Example B
1. Preparation of an Indicator Material for Assessing Body Odor
[0258] Mixtures of 3-hydroxy-3-methylhexanoic acid (Substance (A))
and 3-mercapto-3-methylhexanol (Substance (C)) was prepared at a
0.015% diluted concentration in ethanol, wherein Substance (A)
:Substance (C)=50:1, 100:1 and 200:1, as indicator materials for
assessing body odor for Example B-1, B-2 and B-3 respectively.
Also, a 0.015% diluted solution of 3-methyl-2-hexenoic acid in
ethanol was prepared as an indicator material for assessing body
odor for Comparative Example B-2.
2. Evaluation of a Deodorizing and Masking Effect of an Indicator
Material for Assessing Body Odor Against an Apocrine Odor
[0259] The test subjects were 6 people in the Group A (who were
recognized to have no axillary odor) of Example A. Each of the
above-mentioned indicator materials for assessing body odor
(Examples B-1 to B-3 in Table 2) was sprayed to an axillary region
of each test subjects once (about 30 mg) to apply the indicator
material for assessing body odor to the axillary regions of the
test subjects having no apocrine odor. Then, after each of
commercially available deodorant products A to E was sprayed to an
axillary region of each test subject, to which the indicator
material for assessing body odor was applied, for about 1 second,
an organoleptic test by 10 professional panel members (5 males and
5 females) was conducted concerning a deodorant and masking effect
of the deodorants.
[0260] Also, after each of the commercially available deodorant
products A to E was sprayed to an axillary region of each of the
test subjects who were recognized to have an apocrine odor (6
people who were recognized to have a strong apocrine odor in the
Group C of Example A (Subject identification Nos. 1 to 6)) for
about 1 second, an organoleptic test by 10 professional panel
members (5 males and 5 females) was conducted concerning the
deodorant and masking effect of the deodorants (Comparative example
B-1).
[0261] The organoleptic test was conducted in a room maintained at
25.degree. C. and 65% humidity. The evaluation was regarding 5
commercial products (commercially available deodorant products A to
E shown in Table 2) in total so as to conducted one commercial
product per day. An odor of the axillary region after treated with
each commercial product evaluated according to the following
criteria.
<The Evaluation Criteria for Odor>
[0262] 0: Apocrine odor is not decreased [0263] 1: Apocrine odor is
slightly decreased [0264] 2: Apocrine odor is decreased [0265] 3:
Apocrine odor is significantly decreased
[0266] The deodorant and masking effect of the commercially
available deodorant products was assessed by using organoleptic
test results of odor (average value) by 10 professional panel
members, which was presented in the following 4 levels.
<The Criteria of Effectiveness Evaluation for Deodorants (Marks
in Parentheses are Used in Table 2)>
[0267] 0 or more and less than 0.5: Not effective (X) [0268] 0.5 or
more and less than 1.5: Slightly effective (.DELTA.) [0269] 1.5 or
more and less than 2.5: Effective (.largecircle.) [0270] 2.5 or
more and less than 3.0: Significantly effective (.circleincircle.)
3. Results of Evaluation
[0271] Accuracy of the apocrine odor reproduced by the indicator
material for assessing body odor is proved when the deodorant
effect of the deodorants treated to the test subjects preliminarily
treated by the indicator material for assessing body odor is
considerably consistent with the deodorant effect of the deodorants
treated to the test subjects having actual apocrine odors.
[0272] In Table 2, the deodorant effects of the deodorants treated
to the test subjects preliminarily treated by the indicator
material for assessing body odor of Comparative example B-2 and
Examples B-1 to B-3 are compared based on the deodorant effect of
the deodorants treated to the test subjects having actual apocrine
odors (Comparative example B-1). As a result, Example B-2 was
completely consistent with Comparative example B-1 and Examples B-1
and B-3 were mostly consistent with Comparative example B-1, which
proved that the present invention as an indicator material can
accurately reproduce the apocrine odor, and the deodorant and
masking effect of a deodorant can be accurately assessed by the
indicator material according to the present invention.
[0273] On the contrary, Comparative example B-2 was not consistent
with Comparative B-1 at all, and Comparative example B-2 using
3-methyl-2-hexenoic acid could not assess the deodorant and masking
effect of deodorants as a standard substance, which proved that
Comparative example B-2 can not accurately reproduce the apocrine
odor. TABLE-US-00002 TABLE 2 Assessment results of effectiveness
(deodorant and masking) of deodorants Comparative Example B-1
Example Apocrine odor B-2 B-1 B-2 B-3 naturally 3-methyl-2- Mixture
of 50:1 Mixture of 100:1 Mixture of 200:1 Deodorant generated from
hexenoic (Component A: (Component A: (Component A: product axillary
acid Component C) Component C) Component C) Commercial .DELTA. X
.DELTA. .DELTA. .DELTA. product A Commercial .circleincircle.
.DELTA. .largecircle. .circleincircle. .circleincircle. product B
Commercial X .largecircle. X X .DELTA. product C Commercial .DELTA.
X .DELTA. .DELTA. .largecircle. product D Commercial .largecircle.
.DELTA. .DELTA. .largecircle. .largecircle. product E Consistency
-- Not Slightly Completely Slightly with consistent consistent
consistent consistent Comparative example B-1
Example C
1. Screening of Test Subjects
[0274] 65 Healthy Japanese males cooperated as volunteers. Each of
them wore a new white T-shirt made of cotton for 24 hours
consecutively. After the T-shirts were collected, the strength of
apocrine odor and acid odor was assessed by 7 professional panel
members about axillary regions of T-shirts based on the following
criteria.
<The Criteria of Organoleptic Test>
[0275] Intensity 0: Odorless [0276] Intensity 1: Slight odor [0277]
Intensity 2: Weak odor [0278] Intensity 3: Easily perceptible Odor
[0279] Intensity 4: Strong odor [0280] Intensity 5: Very strong
odor
[0281] As a result, 10 people of test subjects were recognized to
have apocrine odor of intensity 3 or more, 3 people were recognized
to have apocrine odor of intensity 1 or 2, 52 people were
recognized to have apocrine odor of intensity 0.
2. Quantification of 3-hydroxy-3-methyl hexanoic acid
[0282] 4 people who were recognized to have apocrine odor of
intensity 3 or more (Subject identification Nos. A to D), 3 people
who were recognized to have apocrine odor of intensity 1 or 2
(Subject identification Nos. E to G), 3 people who were recognize
to have no apocrine odor (intensity 0) (Subject identification Nos.
H to J) were selected as test subjects.
[0283] A cotton pad was sewn on a part of a T-shirt corresponding
to the axillary region of the sword arm. After the test subjects
wore the T-shirts for 24 hours, the T-shirts were collected and the
cotton pad of the axillary region was subject to the acid-base
extraction to extract acid component originated from perspiration.
Then, by using the silica gel mini column (Varian Bond Elute Jr),
10 mL of diethyl ether, and 10 mL of methanol, the acid extract was
separated in to a diethyl ether fraction and a methanol fraction.
After concentrating the methanol fraction once, 1 mL of diluted
solution was prepared by a measuring flask (manufactured by
DURAN).
[0284] 1 .mu.L of the diluted solution was analyzed by the gas
chromatography-mass spectrometer (GC-MS). Using a synthesized
3-hydroxy-3-methylhexanoic acid as a standard material (standard),
an calibration curve was drawn, and the amount of
3-hydroxy-3-methylhexanoic acid contained in perspiration was
measured.
[0285] The result of organoleptic test on axillary odor and
quantification of 3-hydroxy-3-methylhexanoic acid is shown in FIG.
3. 3-Hydroxy-3-methylhexanoic acid was detected from the methanol
fractions of all test subjects who were recognized to have apocrine
odor (Subject identification Nos. A to G). On the contrary,
3-hydroxy-3-methylhexanoic acid was not detected from the diethyl
ether fraction. TABLE-US-00003 TABLE 3 Organoleptic test Detected
amount of 3-hydroxy- Subject Intensity Intensity 3-methyl hexanoic
acid (.mu.g) identification of apocrine of acid Methanol Ether No.
odor odor fraction fraction A 4 4 33.93 Not Detected B 5 5 64.29 ''
C 3 3 15.71 '' D 4 4 37.5 '' E 2 2 1.71 '' F 1 2 0.87 '' G 2 2 1.39
'' H 0 1 Not Detected '' I 0 2 '' '' J 0 1 '' ''
<The Criteria of Organoleptic Test> [0286] Intensity 0:
Odorless [0287] Intensity 1: Slight odor [0288] Intensity 2: Weak
odor [0289] Intensity 3: Easily perceptible odor [0290] Intensity
4: Strong odor [0291] Intensity 5: Very strong odor
[0292] In the group recognized to have apocrine odor of intensity 3
or more (Subject identification Nos. A to D), the detected amount
of 3-hydroxy-3-methylhexanoic acid was within the scope of 15.71 to
64.29 .mu.g. In the group recognized to have apocrine odor of
intensity 1 or 2 (Subject identification Nos. E to G), the detected
amount of 3-hydroxy-3-methylhexanoic acid was within the scope of
0.87 to 1.71 .mu.g. On the contrary, in the group recognized to
have no apocrine odor (intensity 0) (Subject identification Nos. H
to J), 3-hydroxy-3-methylhexanoic acid was not detected from any
test subjects.
Example D
[0293] The methanol fraction obtained in Example C (1 .mu.L was
used for the GC-MS analysis) was concentrated to 0.5 mL and
transferred to a test tube equipped with a screw cock (NR-10,
manufactured by Maruemu). 100 .mu.L of 20 mM 2-nitrophenylhydrazine
solution prepared by using 40 mM hydrochloric acid-hydrochloric
acid ethanol (3:1, v/v), 100 .mu.L of 3v/v % pyridine in ethanol
solution and 100 .mu.L of 250 mM EDC
(1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) in ethanol solution
were successively added. A blank sample for comparing hue (0.5 mL
of methanol was added) was similarly prepared. After the mixtures
were heated at 60.degree. C. for 20 minutes, 50.mu.L of 15% (W/V)
potassium hydroxide solution prepared by water-methanol (1:4, v/v)
mixture was added followed by heating at 60.degree. C. until the
brown color of the blank disappears (for 15 minutes). After cooling
down to ambient temperature, the colors of reacted solutions were
observed by the naked eye.
[0294] The results are shown in Table 4. The blank sample exhibited
bright yellow color originated from unreacted reagent. On the other
hand, the methanol fractions of the test subjects in which
3-hydroxy-3-methylhexanoic acid was detected exhibited magenta
color due to production of acid hydrazide. The hue changed
proportionally to the detected amount from brownish yellow to dark
magenta. TABLE-US-00004 TABLE 4 Intensity Detected amount Subject
of of 3-hydroxy-3- identification apocrine methyhexanoic Color Hue
of No. odor acid (.mu.g) change test solution A 4 33.93
.smallcircle. Dark magenta B 5 64.29 .smallcircle. Very dark
magenta C 3 15.71 .smallcircle. Slightly dark sienna D 4 37.5
.smallcircle. Dark magenta .smallcircle. Slightly brownish E 2 1.71
yellow .smallcircle. Slightly brownish F 1 0.87 yellow
.smallcircle. Slightly brownish G 2 1.39 yellow H 0 Not Detected x
Light yellow(on a I 0 '' x level with blank) J 0 '' x
<The Criteria of Evaluation> [0295] .largecircle.: The change
of hue was recognized adequately [0296] .DELTA.: The change of hue
was recognized [0297] X: The change of hue was not recognized (on a
level with the blank)
[0298] In the group recognized to have apocrine odor of intensity 3
or more, the color of the reacted solution was within the scope of
red to magenta. In the group recognized to have apocrine odor of
intensity 1 or 2, the color of the reaction solution was brownish
yellow. On the contrary, in the group recognized to have no
apocrine odor (intensity 0), the test solution of all test subjects
were not colored and the hue was on a level with the blank
sample.
[0299] Next, the hue of test solutions was quantified using the
calorimeter. The hue was graphed by selecting the color difference
(.DELTA.E*ab) on the vertical axis, and the organoleptic test
result of apocrine odor in Example C on the horizontal axis. The
result was, as shown in FIG. 11, the color difference (.DELTA.E*ab)
increased proportional to the intensity of apocrine odor.
<Conditions for Measurement by Calorimeter>
[0300] Equipment: Colorimeter CT-310 (manufactured by Minolta Co.,
Ltd.) [0301] Cell optical length: 2 mm [0302] Temperature:
23.degree. C.
[0303] The color difference was obtained from the following
expression using the measured data of the blank sample as the
color-difference standard color (L*.sub.t, a*.sub.t, b*.sub.t) and
the measured data as (L*, a*, b*) .DELTA.E*ab= {square root over
(((.DELTA.L*).sup.2+(.DELTA.a*).sup.2+(.DELTA.b*).sup.2))}
Expression 1 [0304] wherein .DELTA.L*=L*-L.sub.t [0305]
.DELTA.a*=a*-a.sub.t [0306] .DELTA.b*=b*-b.sub.t
Example E
[0307] 1 mL of the diethyl ether fraction obtained in Example C was
transferred to a test tube equipped with a screw cock (NR-10,
manufactured by Maruemu). After the solvent was removed in a water
bath of 45.degree. C. and 0.5 mL of methanol was added, the test
solutions as well as a blank sample for comparing hue (0.5 mL of
methanol was added) were subject to the coloration test in the
manner similar to Example D using a coloration reagent. Together
with the methanol fraction obtained in Example D, the hue was
quantified by means of an ultraviolet-visible spectrometer. The
data was graphed by selecting the absorbance of the methanol
fraction on the vertical axis, and the absorbance of the ether
fraction on the horizontal axis to evaluate a level of contribution
of apocrine odor and acid odor to each test subject.
[0308] The result was, as shown in FIG. 12, a person whose methanol
fraction of the acid extract exhibits strong color, i.e. a person
recognized to have apocrine odor, tends to have the diethyl ether
fraction which exhibits strong coloration (absorption). On the
contrary, a person recognized to have no apocrine odor, i.e. a
person whose methanol fraction does not exhibit color tends to have
the diethyl ether fraction which exhibits weak coloration
(absorption).
<Conditions for Measurement by Ultraviolet-Visible
Spectrometer>
[0309] Equipment: BECHMAN DU-600 [0310] Cell optical length: 10 mm
[0311] Temperature: 23.degree. C. [0312] Measurement method of
data: Peak area was measured by deducting the absorption spectrum
of the blank sample from the absorption spectrum of a sample at 530
nm.
Example F
[0313] 10 Test subjects of Example C were test subjects for Example
F. The methanol fraction of acid extract of perspiration separated
in the manner similar to Example C was concentrated to 0.5 mL, and
thereafter transferred to a test tube. Together with a blank sample
(0.5 mL of methanol was added) for comparing hue, 25 uL of 1.0%
9-anthryldiazomethane in acetone solution was added to each sample.
The each sample was sealed and left for about 1 hours at ambient
temperature. Then, using an ultraviolet lamp (manufactured by
Ultraviolet Corporation, long wavelength type of 365 nm), the
emitted fluorescence was observed by the naked eye. A result is
shown in Table 5. TABLE-US-00005 TABLE 5 Subject identification
Intensity of Color Fluorescence of test No. apocrine odor change
solution A 4 .smallcircle. Very strong B 5 .smallcircle. Very
strong C 3 .smallcircle. Slightly strong D 4 .smallcircle. Very
strong E 2 .smallcircle. Strong F 1 .smallcircle. Strong G 2
.smallcircle. Strong H 0 x Weak(on a level with blank) I 0 x J 0
x
[0314] As a result shown in Table 5, the samples of test subjects
recognized to have apocrine odor in Example C emitted stronger
fluorescence proportional to the intensity of the apocrine odor. On
the contrary, a strong fluorescence was not observed regarding the
blank sample.
* * * * *