U.S. patent application number 13/058844 was filed with the patent office on 2011-06-16 for polymer for filler for preprocessing column.
This patent application is currently assigned to WAKO PURE CHEMICAL INDUSTRIES, LTD.. Invention is credited to Yoshinori Inoue, Mamoru Kubota, Hitoshi Uemori, Kimiko Yoshida.
Application Number | 20110138935 13/058844 |
Document ID | / |
Family ID | 41668954 |
Filed Date | 2011-06-16 |
United States Patent
Application |
20110138935 |
Kind Code |
A1 |
Inoue; Yoshinori ; et
al. |
June 16, 2011 |
POLYMER FOR FILLER FOR PREPROCESSING COLUMN
Abstract
It is a subject of the present invention to provide a novel
polymer; a filler for measuring a perfluoro compound having an
acidic group at terminal which comprises said polymer; a column
filled with said filler; and a measuring method for a perfluoro
compound having an acidic group at terminal by using said column;
as well as a filler for measuring a drug comprising the
above-described polymer; a column filled with said filler; and a
measuring method for a drug by using said column. The present
invention relates to a polymer obtained by introducing an anion
exchanging group to a glycidyl group in a polymer obtained by
polymerizing the compound represented by the following general
formula [1]: ##STR00001## and the compound represented by the
following general formula [2]: ##STR00002## a filler (column) for
measuring an acidic group binding perfluoro compound at terminal
which comprises said polymer; a method for measuring said perfluoro
compound by using said column; a filler (column) for measuring a
drug which comprises the above-described polymer; and a method for
measuring a drug by using said column.
Inventors: |
Inoue; Yoshinori; (Tokyo,
JP) ; Yoshida; Kimiko; (Hyogo, JP) ; Kubota;
Mamoru; (Hyogo, JP) ; Uemori; Hitoshi; (Hyogo,
JP) |
Assignee: |
WAKO PURE CHEMICAL INDUSTRIES,
LTD.
Osaka
JP
NIPPON FILCON CO., LTD.
Tokyo
JP
|
Family ID: |
41668954 |
Appl. No.: |
13/058844 |
Filed: |
August 10, 2009 |
PCT Filed: |
August 10, 2009 |
PCT NO: |
PCT/JP2009/064128 |
371 Date: |
February 11, 2011 |
Current U.S.
Class: |
73/863.21 ;
521/31; 521/32 |
Current CPC
Class: |
Y10T 436/201666
20150115; Y10T 436/173845 20150115; Y10T 436/182 20150115; C08F
220/36 20130101; C08F 8/32 20130101; C08F 236/20 20130101; B01J
41/14 20130101; C08F 8/30 20130101; B01J 41/20 20130101; Y10T
436/145555 20150115; Y10T 436/141111 20150115; B01J 2220/54
20130101; Y10T 436/193333 20150115; C08F 8/30 20130101; C08F 8/32
20130101; G01N 30/02 20130101; G01N 30/02 20130101; Y10T 436/147777
20150115; C08F 224/00 20130101; C08F 236/22 20130101; G01N 30/96
20130101; C08F 220/32 20130101; C08F 220/32 20130101; C08F 220/32
20130101; B01D 15/363 20130101 |
Class at
Publication: |
73/863.21 ;
521/31; 521/32 |
International
Class: |
G01N 1/10 20060101
G01N001/10; B01J 41/14 20060101 B01J041/14 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 12, 2008 |
JP |
2008-207879 |
Nov 27, 2008 |
JP |
2008-301822 |
Claims
1. A polymer obtained by introducing an anion exchanging group to a
glycidyl group of a polymer obtained by polymerizing a compound
represented by the following general formula [1]: ##STR00036##
(wherein each of R.sub.1 and R.sub.2 independently represents a
hydrogen atom, a linear alkyl group having 1 to 3 carbon atoms or a
halogen atom, Bz represents benzene ring), and a compound
represented by the following general formula [2]: ##STR00037##
(wherein R.sub.9 represents a hydrogen atom, a linear alkyl group
having 1 to 3 carbon atoms or a halogen atom, R.sub.10 represents a
linear alkylene group having 1 to 3 carbon atoms, each of R.sub.11
and R.sub.12 independently represents a hydrogen atom or a linear
alkyl group having 1 to 6 carbon atoms).
2. The polymer according to claim 1, wherein the anion exchanging
group is a tertiary amino group.
3. The polymer according to claim 1, wherein the anion exchanging
group is a morpholino group or a diethylamino group.
4. The polymer according to claim 1, wherein the compound
represented by the general formula [1] is divinylbenzene.
5. The polymer according to claim 1, wherein the compound
represented by the general formula [2] is the compound represented
by the following general formula [2']: ##STR00038##
6. A filler for measuring a perfluoro compound having an acidic
group at terminal, comprising the polymer according to claim 1.
7. The filler according to claim 6, wherein the acidic group at
terminal of the perfluoro compound is a carboxyl group or a sulfo
group.
8. A preprocessing column for measuring a perfluoro compound having
the acidic group at terminal which is filled with the filler
according to claim 6.
9. A measuring method for measuring a perfluoro compound having an
acidic group at terminal, characterized in using the column
according to claim 8.
10. A measuring method for a perfluoro compound comprising a step
for adsorbing said perfluoro compound by passing the sample
containing said perfluoro compound having an acidic group at
terminal to the column according to claim 8; a step for eluting
said perfluoro compound by eluate; and a step for measuring the
obtained perfluoro compound.
11. The polymer according to claim 1, wherein the anion exchanging
group is an ammonium group represented by the following general
formula [3]: ##STR00039## (wherein each of R.sub.3 represents
independently a linear alkyl group having 1 to 3 carbon atoms,
R.sub.4 represents a linear alkyl group having 1 to 6 carbon
atoms).
12. The polymer according to claim 11, wherein R.sub.3 represents a
methyl group and R.sub.4 represents an ethyl group in the general
formula [3].
13. The polymer according to claim 11, wherein said polymer is
obtained by introducing the ammonium group represented by the
general formula [3] to the glycidyl group in the polymer obtained
by polymerizing monomers containing the compound represented by the
general formula [1] and the compound represented by the general
formula [2] and further the compound represented by the following
general formula [4]: ##STR00040## (wherein each of three R.sub.15
represents independently a linear alkylene group having 1 to 3
carbon atoms, each of three R.sub.16 independently represents a
hydrogen atom, a linear alkyl group having 1 to 3 carbon atoms or a
halogen atom).
14. The polymer according to claim 13, wherein R.sub.15 in the
general formula [4] is a methylene group and R.sub.16 is a hydrogen
atom.
15. The polymer according to claim 11, wherein the compound
represented by the general formula [1] is divinylbenzene.
16. The polymer according to claim 11, wherein the compound
represented by the general formula [2] is the compound represented
by the following general formula [2']: ##STR00041##
17. A filler for measuring drugs which comprises the polymer
according to claim 11.
18. A preprocessing column filled with the filler according to
claim 17.
19. A measuring method of a drug characterized in using the column
according to claim 18.
20. A measuring method of a drug comprising a step for adsorbing
said drug by passing the sample containing said drug to the column
according to claim 18; a step for eluting said drug by eluate; and
a step for measuring the eluted drug.
Description
TECHNICAL FIELD
[0001] The present invention relates to a novel polymer; a filler
for measuring a perfluoro compound having an acidic group at
terminal which comprises said polymer; a column filled with said
filler; and a measuring method for a perfluoro compound having an
acidic group at terminal by using said column; as well as a filler
for measuring a drug which comprises the above-described polymer; a
column filled with said filler; and a measuring method for a drug
by using said column.
BACKGROUND ART
[0002] Perfluoro compounds having acidic groups at terminal such as
perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid
(PFOA), which are perfluorinated fluorine compounds, have been
widely used for various industrial products such as household
articles, construction materials, semiconductors because of having
excellent properties of heat resistance, chemical resistance,
weathering resistance and the like. However, it has been recently
found that such perfluoro compounds containing acidic group have
remained in water of river, lake, seawater etc., and have been
accumulating in the body of human and living organism because these
compounds tend to migrate into water environment due to
water-solubility, and are chemically very stable and persistent.
Therefore, these compounds have been attracting the attention as
the items to be monitored of the global environmental pollution,
and the nationwide research on the environmental pollution by
perfluoro compounds having acidic groups at terminal has been
carried out. And as a micro-analytical method for perfluoro
compound contained in the environmental water in this research, the
method for analyzing the samples pretreated by using a solid-phase
extraction column with LC/MS/MS and the like is employed. For
example, the method for analyzing perfluoro compounds having acidic
groups at terminal by using the solid-phase extraction method used
with a processing column is described in non-patent reference 1 and
the like. However, 10 to 20 kinds of perfluoro compounds having
acidic groups at terminal are present depending on the difference
of acidic groups at terminal and the number of carbon atoms
thereof, and we have confirmed that all perfluoro compounds having
acidic groups at terminal cannot be simultaneously analyzed by
using the measuring method being carried out in the above-described
reference. Therefore, the development of the method by which
perfluoro compounds having acidic groups at terminal can be
simultaneously and efficiently analyzed, has been desired.
[0003] On the other hand, need of the test of medicaments (drugs)
contained in biological samples is recently increasing. In
particular, doping test in the sports world needs to inspect
various drugs, and needs to bring about no inspection mistake even
when it is a trace amount. Usually, this drug test is carried out
by solid-phase extracting with the preprocessing column, and
further analyzing and detecting the obtained extraction liquid by
using high-performance liquid chromatography or gas chromatography.
However, since the above-described drug test is carried out by
targeting the various drugs, the development of the preprocessing
column which can adsorb any type of the basic, neutral, or acidic
compounds, has been desired. On the other hand, patent reference 1
describes the porous resin compound having an ion-exchanging group,
a hydrophobic component and a hydrophilic component as a column
(filler) having such a wide adsorption target. However, even these
compounds, they cannot efficiently adsorb all of the
above-described target components. Therefore, further improvement
has been needed.
[Prior Art Documents]
[Patent Documents]
[0004] Patent Reference 1: JP-A-2002-517574;
[Non-Patent Documents]
[0005] NON-Patent Reference 1: Journal of Chromatography A,
19093(2005)89-97;
SUMMARY OF INVENTION
Problem to be Solved by the Invention
[0006] Considering the above-described situation, the present
inventors have extensively studied a way to attain the
above-described object, and consequently, have found that a polymer
obtained by introducing an anion-exchanging group into a glycidyl
group in the polymer obtained by polymerization of the compound
represented by the following general formula [1]:
##STR00003##
(wherein each of R.sub.1 and R.sub.2 independently represents a
hydrogen atom, a linear alkyl group having 1 to 3 carbon atoms or a
halogen atom, Bz represents benzene ring) and the compound
represented by following general formula [2]:
##STR00004##
(wherein R.sub.9 represents a hydrogen atom, a linear alkyl group
having 1 to 3 carbon atoms or a halogen atom, R.sub.10 represents a
linear alkylene group having 1 to 3 carbon atoms, each of R.sub.11
and R.sub.12 independently represents a hydrogen atom, or a linear
alkyl group having 1 to 6 carbon atoms) can become a filler for
solving the above-described problems. Then, they have completed the
present invention. That is, it has been found that the filler
comprising said polymer can efficiently adsorb the various
perfluoro compounds which have the acidic group at terminal
(hereinafter, this compound may be abbreviated as the perfluoro
compound having the acidic group), also, the filler comprising the
above-described polymer can efficiently adsorb any one of the
basic, neutral, or/and acidic compound. Thus, the present invention
has been completed. It is a subject of the present invention to
provide a novel polymer; a filler which can efficiently adsorb
various acidic group binding perfluoro compound; and a measuring
method for the perfluoro compounds having the acidic group by using
said filler; as well as a filler which can efficiently adsorb the
basic, neutral or/and acidic drugs; and a measuring method for the
drugs by using said filler.
Means for Solving the Problems
[0007] The present invention relates to:
[0008] (1) a polymer obtained by introducing an anion exchanging
group into a glycidyl group in the polymer obtained by
polymerization of the compound represented by the following general
formula [1]:
##STR00005##
(wherein, each of R.sub.1 and R.sub.2 independently represents a
hydrogen atom, a linear alkyl group having 1 to 3 carbon atoms or a
halogen atom, Bz represents benzene ring) and the compound
represented by the following general formula [2]:
##STR00006##
(wherein, R.sub.9 represents a hydrogen atom, a linear alkyl group
having 1 to 3 carbon atoms or a halogen atom, R.sub.10 represents a
linear alkylene group having 1 to 3 carbon atoms, each of R.sub.11
and R.sub.12 independently represents a hydrogen atom, or a linear
alkyl group having 1 to 6 carbon atoms);
[0009] (2) a filler for measuring a perfluoro compound having an
acidic group at terminal, comprising said polymer;
[0010] (3) a preprocessing column filled with said filler for
measuring a perfluoro compound having an acidic group at
terminal;
[0011] (4) a measuring method for a perfluoro compound having an
acidic group at terminal characterized in using said column;
[0012] (5) a filler comprising the above-described polymer for
measuring a drug;
[0013] (6) a preprocessing column filled with said filler for
measuring a drug; and
[0014] (7) a measuring method for a drug characterized in using
said column.
Effect of the Invention
[0015] The filler comprising the polymer of the present invention
can efficiently adsorb various perfluoro compounds having the
acidic group. Therefore, by the method using said filler,
particularly, by the measuring method in which solid-phase
extraction is carried out by using said filler, it is possible to
measure simultaneously and efficiently various perfluoro compounds
having the acidic group. Further, by using LC/MS/MS as the
separation measurement, it is possible to measure sensitively.
[0016] In addition, the filler comprising the polymer of the
present invention can adsorb the target substances of the basic,
neutral or/and acidic compound and the like equally or more
efficiently comparing with the conventional filler for the
preprocessing column having broad scope. Therefore, a plurality of
targeting substances can be simultaneously and efficiently
separated by concentration by means of one time pretreatment.
Particularly, when drugs are provided as the target substances, the
column is very useful as the preprocessing column for measuring the
drugs because of capability of efficient adsorption. Furthermore,
the adsorbed basic, neutral, or/and acidic compounds can be
separately eluted as basic and neutral compounds and acidic
compounds. Therefore, this method also gives the effect that
separation analysis becomes easy for the many target
substances.
Modes for Carrying Out the Invention
[0017] Alkyl groups having 1 to 3 carbon atoms in R.sub.1 and
R.sub.2 of the general formula [1] include, specifically, for
example, a methyl group, an ethyl group, an n-propyl group and the
like, and a methyl group is preferable.
[0018] Halogen atoms in R.sub.1 and R.sub.2 of the general formula
[1] include a fluorine atom, a chlorine atom, a bromine atom, an
iodine atom and the like.
[0019] The compounds represented by the general formula [1]
include, specifically, for example,
##STR00007##
and the like.
##STR00008##
are preferable, and among them,
##STR00009##
is particularly preferable.
[0020] Linear alkyl groups having 1 to 3 carbon atoms in R.sub.9 of
the general formula [2] include, specifically, for example, a
methyl group, an ethyl group, an n-propyl group and the like, and a
methyl group is preferable.
[0021] Halogen atoms in R.sub.9 of the general formula [2] include
a fluorine atom, a chlorine atom, a bromine atom, an iodine atom
and the like.
[0022] Linear alkylene groups having 1 to 3 carbon atoms in
R.sub.10 of the general formula [2] include, for example, a
methylene group, an ethylene group, an n-propylene group and the
like, and a methylene group is preferable.
[0023] Linear alkyl groups having 1 to 6 carbon atoms in R.sub.11
and R.sub.12 of the general formula [2] include, for example, a
methyl group, an ethyl group, an n-propyl group, an n-butyl group,
an n-pentyl group, an n-hexyl group and the like. Among them, a
methyl group is preferable.
[0024] Specific examples of the general formula [2] include, for
example,
##STR00010##
and the like.
##STR00011##
and the like are preferable. Among them,
##STR00012##
is particularly preferable.
[0025] Anion exchanging groups relevant to the present invention
include, for example, a primary amino group, a secondary amino
group, a tertiary amino group, a quaternary ammonium group and the
like. Secondary amino groups include, for example, the group
represented by --NHR.sub.20 (R.sub.20 represents an alkyl group
which may contain an oxygen atom in the chain), and tertiary amino
groups include the group represented by --NHR.sub.21R.sub.21' (each
of R.sub.21, R.sub.21' independently represents an alkyl group
which may contain oxygen atoms in the chain) or a cyclic amino
group, and quaternary ammonium groups include the group represented
by the following formula [3]:
##STR00013##
(wherein each of R.sub.3 independently represents a linear alkyl
group having 1 to 3 carbon atoms, and R.sub.4 represents a linear
alkyl group having 1 to 6 carbon atoms). As said anion exchanging
group, a tertiary amino group is preferable when polymer is used as
the filler for measuring the perfluoro compound having the acidic
group, and a quaternary ammonium group is preferable when polymer
is used as the filler for measuring the drugs.
[0026] Alkyl groups in the alkyl group which may have oxygen atoms
represented by R.sub.20 in the above-described secondary amino
group may be linear, branched, or cyclic group, and a linear group
is preferable. Alkyl groups include the group having normally 1 to
6 carbon atoms, preferably 1 to 4 carbon atoms, and more preferably
1 or 2 carbon atoms, and are specifically represented by, for
example, a methyl group, an ethyl group, an n-propyl group, an
isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl
group, a tert-butyl group, an n-pentyl group, an isopentyl group, a
sec-pentyl group, a tert-pentyl group, a neopentyl group, an
n-hexyl group, an isohexyl group, a sec-hexyl group, a tert-hexyl
group, an n-heptyl group, an isoheptyl group, a sec-heptyl group, a
tert-heptyl group, an n-octyl group, a sec-octyl group, a
tert-octyl group, a nonyl group, a decyl group, an undecyl group, a
dodecyl group, a cyclopropyl group, a cyclopentyl group, a
cyclohexyl group, a cycloheptyl group, a cyclooctyl group, a
cyclononyl group, a cyclodecyl group, a cycloundecyl group, a
cyclododecyl group and the like, and a methyl group and an ethyl
group are preferable, and an ethyl group is more preferable. In
addition, alkyl groups having oxygen atoms include the alkyl group
having normally 1 to 5, preferably 1 to 2, more preferably 1 oxygen
atom in the chain of the above-described alkyl group having 2 to 6
carbon atoms, and specific examples include, for example,
--CH.sub.2--O--CH.sub.3, --CH(CH.sub.3)--O--CH.sub.3,
--CH.sub.2--CH(CH.sub.3)--O--CH.sub.3,
--CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2--O--C(CH.sub.3).sub.3,
--CH.sub.2CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.2--O--CH(CH.sub.3)CH.sub.3,
--CH.sub.2CH(CH.sub.3)--O--CH.sub.2CH(CH.sub.3)CH.sub.3,
--CH.sub.2--O--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2--O--CH.sub.3,
--CH.sub.2CH.sub.2--O--CH(CH.sub.2CH.sub.3)CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2CH.sub.2CH-
.sub.2CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--O--C(CH.sub.3).sub.3,
--CH.sub.2CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2CH.sub.2--O--CH.sub.2CH.su-
b.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.-
2CH.sub.2CH.sub.2--O--CH.sub.3,
--CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2--O--CH.sub.2-
CH.sub.3,
--CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2--O--
-CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2--O--CH.sub.2CH.sub.2--O--CH.sub.2CH(CH.sub.3)
and the like.
[0027] Preferable specific examples of the above-described
secondary amino group include, for example, a methylamino group, an
ethylamino group, an n-propylamino group and the like.
[0028] Alkyl groups which may contain oxygen atoms represented by
R.sub.21, or R.sub.21' in the above-described tertiary amino group
include the same as the alkyl groups which may contain oxygen atoms
represented by the above-described R.sub.20, and include, as
preferable groups, a methyl group, an ethyl group and an n-propyl
group, and among them, an ethyl group is preferable. It should be
noted that, it is more preferable that both of R.sub.21 and
R.sub.21' are the same groups. Specific examples of tertiary amino
groups represented by --NR.sub.21 or R.sub.21' include, for
example, a dimethylamino group, a diethylamino group, a
methylethylamino group, a di-n-propylamino group, a
diisopropylamino group, an 1-phenylethylamino group and the like,
and among them, a diethylamino group is preferable.
[0029] Cyclic amino groups of tertiary amino group include an amino
group constituting normally 3 to 12 membered ring, preferably 5 to
6 membered ring in cyclic part, specifically include, for example,
the groups derived from membered rings such as an ethyleneimino
group, for example, 5 membered rings such as a pyrrolidino group,
imidazolino group, a pyrazolidino group, for example, 6 membered
rings such as a piperidino group, a morpholino group, a piperazino
group, for example, the groups derived from bicyclo rings such as a
quinuclidino group, and among them, 6 membered rings such as a
piperidino group, a morpholino group, a piperazino group are
preferable, and further, a morpholino group is preferable.
[0030] Linear alkyl groups having 1 to 3 carbon atoms in R.sub.3 of
quaternary ammonium groups represented by the general formula [3]
include, specifically, for example, a methyl group, an ethyl group,
an n-propyl group and the like, and a methyl group is
preferable.
[0031] Linear alkyl groups having 1 to 6 carbon atoms in R.sub.4 of
quaternary ammonium groups represented by the general formula [3]
include, for example, a methyl group, an ethyl group, an n-propyl
group, an n-butyl group, an n-pentyl group, an n-hexyl group and
the like, and among them, a methyl group is preferable.
[0032] Ammonium groups represented by the general formula [3]
include, specifically, for example, a trimethylammonium group, an
ethyldimethylammonium group, a dimethyl-n-propylammonium group, an
n-butyldimethylammonium group, a dimethyl-n-pentylammonium group, a
dimethyl-n-hexylammonium group, a diethylmethylammonium group, a
triethylammonium group, a tri-n-ammonium group and the like, and
among them, a trimethylammonium group, an ethyldimethylammonium
group are preferable, and an ethyldimethylammonium group is
particularly preferable.
[0033] Linear alkylene groups having 1 to 3 carbon atoms in
R.sub.15 of the general formula [4] include, for example, a
methylene group, an ethylene group, an n-propylene group and the
like, and a methylene group is preferable.
[0034] Linear alkyl groups having 1 to 3 carbon atoms in R.sub.16
of the general formula [4] include, for example, a methyl group, an
ethyl group, an n-propyl group and the like, and a methyl group is
preferable. In addition, halogen atoms in R.sub.16 include a
fluorine atom, a chlorine atom, a bromine atom, an iodine atom and
the like.
[0035] Compounds represented by the general formula [4] include,
specifically, for example,
##STR00014##
and the like, and among them,
##STR00015##
is preferable.
[0036] The polymers obtained by introducing the anion exchanging
group into glycidyl group of the polymer (hereinafter, this polymer
may be abbreviated as the polymer of the present invention)
obtained by polymerization of the compound represented by the
general formula [1] and the compound represented by the general
formula [2] (hereinafter, this polymer may be abbreviated as the
prepolymer relevant to the present invention) include, for example,
the following general formula [3]:
##STR00016##
(wherein R.sub.1, R.sub.2, R.sub.9, R.sub.10, R.sub.11 and R.sub.12
are as above-described. A represents the anion exchanging group
relevant to the present invention), specifically, for example,
##STR00017## ##STR00018## ##STR00019## ##STR00020##
and the like. When the polymer is used as the filler for measuring
the perfluoro compound having the acidic group,
##STR00021##
and the like are preferable. When the polymer is used as the filler
for measuring drugs,
##STR00022##
and the like are preferable.
[0037] In addition, the polymers of the present invention also
include the polymer obtained by introducing an ammonium group
represented by the general formula [3] into a glycidyl group of the
polymer obtained by polymerization of the monomers containing the
compound represented by the general formula [1] and the compound
represented by the general formula [2], and further the compound
represented by the general formula [4], and are specifically
represented by, for example, the following general formula [6]:
##STR00023##
(wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.9, R.sub.10,
R.sub.11, R.sub.12, R.sub.15 and R.sub.16 are as above-described).
Said polymer is preferable for the filler for measuring the drug.
Specific examples of said general formula [6] are, for example,
##STR00024## ##STR00025##
and the like, and among them,
##STR00026##
is preferable.
[0038] Average particle diameter of the polymer of the present
invention is normally 10 to 100 .mu.m, preferably 20 to 100 .mu.m,
more preferably 30 to 70 .mu.m, and micropore diameter is normally
3 to 15 nm. When polymer is used as the filler for measuring the
perfluoro compound having the acidic group, 10 to 15 nm is
preferable. When the polymer is used as the filler for measuring
the drugs, 3 to 5 nm is preferable. Specific surface area of the
polymer is normally 200 to 1000 m.sup.2/g, preferably 300 to 700
m.sup.2/g. When polymer is used as the filler for measuring the
perfluoro compound having the acidic group, 600 to 700 m.sup.2/g is
particularly preferable. When polymer is used as the filler for
measuring drugs, 300 to 500 m.sup.2/g is particularly preferable.
When the polymer of the present invention is synthesized from the
monomers containing the compound represented by the general formula
[1] and the compound represented by the general formula [2], molar
ratio of the compound represented by the general formula [1] and
the compound represented by the general formula [2] is normally 60
to 90:10 to 40, preferable range is 70 to 85:15 to 30. In addition,
when the polymer of the present invention is synthesized from the
monomers containing the compound represented by the general formula
[1], the compound represented by the general formula [2] and the
compound represented by the general formula [4], molar ratio of the
compound represented by the general formula [1], the compound
represented by the general formula [2] and the compound represented
by the general formula [4] is normally 60 to 85:14 to 30:1 to 10,
preferable range is 70 to 80:18 to 25:2 to 5. The polymer of the
present invention is the one in which anion exchanging group is
contained normally 0.1 to 1.0 mmol in 1 g of said polymer,
preferable range is 0.2 to 0.6 mmol.
[0039] Synthetic method of the present invention may be carried out
by polymerizing the monomers containing the compound represented by
the general formula [1] and the compound represented by the general
formula [2], or the monomers (hereinafter, these monomers may be
abbreviated as the monomer group relevant to the present invention)
containing the compound represented by the general formula [1] and
the compound represented by the general formula [2] and the
compound represented by the general formula [4] by the known
polymerization method, and by introducing the anion exchanging
group into glycidyl group at terminal of the obtained polymer
(hereinafter, the polymer may be abbreviated as prepolymer relevant
to the present invention) by the known method (for example,
amination reaction and the like).
[0040] Specifically, for example, after the monomer group relevant
to the present invention and polymerization initiator are dissolved
in the organic solvent which can dissolve these monomer group and
polymerization initiator but is insoluble in water, suspension
polymerization is carried out in the water by adding said organic
solvent to water, and then, the prepolymer relevant to the present
invention is obtained. When the monomers having the compound
represented by the general formula [1] and the compound represented
by the general formula [2] are used as the monomer group relevant
to the present invention, molar ratio of the compound represented
by the general formula [1] and the compound represented by the
general formula [2] is normally 60 to 90:10 to 40, preferable range
is 70 to 85:15 to 30. In addition, when the monomers containing the
compound represented by the general formula [1] and the compound
represented by the general formula [2] and the compound represented
by the general formula [4] are used as the monomer group relevant
to the present invention, molar ratio of the compound represented
by the general formula [1] and the compound represented by the
general formula [2] and the compound represented by the general
formula [4] is normally 60 to 85:14 to 30:1 to 10, preferable range
is to 80:18 to 25:2 to 5. As a polymerization initiator, any one
can be used as long as it is well-known itself, and includes, for
example, azoisobutyronitrile,
2,2'-azobis(2,4-dimethylvaleronitrile), 2,2'-azobis(methyl
2-methypropionate), 2,2'-azobis(2-methylbutyronitrile), benzoyl
peroxide, lauroyl peroxide and the like. Amount of use of the
polymerization initiator is 0.1 to 3% by weight relative to total
weight of the monomer group relevant to the present invention. In
addition, organic solvents to be used here include, for example,
toluene, ethylbenzene, butyl acetate, amyl acetate, octylalcohol,
dodecylalcohol, octane, dodecane and the like. Amount of use of the
solvent is 0.5 to 2 times weight of total weight of monomers
relevant to the present invention. Amount of water to be used here
is normally 1 to 10 times weight relative to sum of the weight of
monomer group relevant to the present invention and the weight of
the above-described organic solvent. In addition, aqueous organic
polymer such as polyvinyl alcohol and methylcellulose as a
suspension stabilizer known in itself are preferably dissolved in
said water, and amount of stabilizer is normally 0.01 to 1% by
weight in the water. Suspension polymerization reaction is carried
out normally at 60 to 90.degree. C., for 4 to 20 hours. After the
reaction, the prepolymer relevant to the present invention can be
obtained by treating according to common procedure of obtaining the
polymer. The obtained prepolymer relevant to the present invention
is preferably classified by using sieves in order to obtain the
prepolymer having the particle diameter suitable for filler, and
also, to obtain the prepolymer having uniform largeness for
performing the excellent capability of adsorption. Particle
diameter of the prepolymer relevant to the present invention
obtained by sieve classification is normally 10 to 100 .mu.m,
preferable range is 20 to 100 .mu.m, more preferable range is 30 to
70 .mu.m. In addition, operation of said classification may be
carried out after obtaining the polymer of the present
invention.
[0041] Subsequently, the obtained prepolymer relevant to the
present invention is suspended or swelled in suitable organic
solvent or the mixed aqueous solution of said organic solvent and
water, and the compound which contains the anion exchanging group
(the anion exchanging group containing compound) is added to said
solution to perform amination reaction, and thus, the polymer of
the present invention can be obtained. The above-described organic
solvents include 1,4-dioxane, tetrahydrofuran, isopropanol,
N,N-dimethylformamide, dimethylsulfoxide and the like. When mixed
aqueous solution of organic solvent and water is used, amount of
water is normally 10 to 60% as an amount in the mixed aqueous
solution, and preferable range is 20 to 50%. The amount of the
above-described organic solvent or mixed aqueous solution of
organic solvent and water is 1 to 10 times weight relative to the
weight of the prepolymer relevant to the present invention. If the
anion exchanging group containing compound is the one which
contains the above-described anion exchanging group, and by which
the desired polymer can be obtained, it is not particularly limited
thereto. For example, and the polymer in which hydrogen atom or
methyl group and the like is bounded to terminal of the
above-described anion exchanging group is exemplified, and amount
of use of the polymer is normally 0.001 to 0.05 mole relative to 1
kg of prepolymer relevant to the present invention, preferable
range is 0.002 to 0.025 mole. Amination reaction is carried out at
room temperature to 60.degree. C., normally for 4 to 30 hours,
preferably for 10 to 20 hours, while stirring if necessary. The
polymer of the present invention is obtained by this reaction.
[0042] The case of employing divinylbenzene as a compound
represented by the general formula [1], glycidyl methacrylate as a
compound represented by the general formula [2], and diethylamine
as the anion exchanging group containing compound, is taken for an
example, and the synthetic method of the polymer of the present
invention is described below. It should be noted that, said polymer
is preferable as the filler for measuring the perfluoro compound
having the acidic group.
[0043] That is, for example, in 100 g of organic solvent such as
toluene, 75 to 90 g of divinylbenzene (0.2 to 0.6 mol as an amount
of pure divinylbenzene), and 10 to 25 g of glycidyl methacrylate
(0.05 to 0.2 mol) and 1 to 3 g of polymerization initiator such as
azobisisobutyronitrile are mixed, the resultant organic solvent is
suspended in 500 to 1000 mL of 0.01 to 1.0% by weight of methyl
cellulose aqueous solution, and is reacted at 60 to 80.degree. C.
for 5 to 10 hours. It should be noted that, when divinyl benzene is
employed, there exists divinyl benzene containing impurities such
as ethylvinylbenzene in commercially available divinylbenzene,
however, in the present invention, the divinylbenzene containing
such impurities may be provided to synthetic reaction as it is. In
that case, although the content of divinylbenzene is reduced, the
divinylbenzene can be used even when molar ratio to the other
compounds is within the range defined in the section of the
above-described polymer of the present inventions. Hereinafter, use
of divinylbenzene is the same as above. By this method, the
prepolymer relevant to the present invention can be obtained. After
the reaction, the resultant polymer is washed with water or
methanol if necessary. Subsequently, 10 g of prepolymer relevant to
the present invention is suspended in 50 to 100 mL of 20 to 50% by
volume of 2-propanol aqueous solution, and 10 to 50 g of
diethylamine (0.1 to 0.5 mol) is added, and further the reaction
solution is reacted at 40 to 60.degree. C. for 10 to 20 hours while
stirring, and then, the polymer of the present invention can be
obtained.
[0044] The case of employing divinylbenzene as a compound
represented by the general formula [1], glycidyl methacrylate as a
compound represented by the general formula [2], and
N,N-dimethylaminoethane as the anion exchanging group containing
compound is taken for an example, and synthetic method of the
polymer of the present invention is described below. It should be
noted that, said polymer is preferable as the filler for measuring
the drugs.
[0045] That is, for example, in 100 g of organic solvent such as
toluene, 75 to 90 g of divinylbenzene (0.2 to 0.6 mol as an amount
of pure divinylbenzene), and 10 to 25 g of glycidyl methacrylate
(0.05 to 0.2 mol) and 1 to 3 g of polymerization initiator such as
azobisisobutyronitrile are mixed, and the resultant organic solvent
is suspended in 500 to 1000 mL of 0.01 to 1.0% by weight of methyl
cellulose aqueous solution, and is reacted at 60 to 80.degree. C.
for 5 to 10 hours. By this method, the prepolymer relevant to the
present invention can be obtained. After the reaction, the
resultant polymer relevant to the present invention is washed with
water or methanol if necessary. Subsequently, 10 g of prepolymer
relevant to the present invention is suspended in 50 to 100 mL of
20 to 50% by volume of 2-propanol aqueous solution, 10 to 50 g of
N,N-dimethylaminoethane (0.1 to 0.5 mol) is added, and the organic
solution is reacted at 40 to 60.degree. C. for 10 to 20 hours while
stirring. Then, the polymer of the present invention can be
obtained.
[0046] The case of employing divinylbenzene as a compound
represented by the general formula [1], glycidyl methacrylate as a
compound represented by the general formula [2], triallyl
isocyanurate as a compound represented by the general formula [4],
and N,N-dimethylaminoethane as the anion exchanging group
containing compound is taken for an example, and synthetic method
of the polymer of the present invention is described below. It
should be noted that, said polymer is preferable as the filler for
measuring the drugs.
[0047] That is, for example, in 100 g of organic solvent such as
toluene, 85 to 70 g of divinylbenzene (0.2 to 0.6 mol as an amount
of pure divinylbenzene), and 10 to 20 g of glycidyl methacrylate
(0.05 to 0.2 mol) and 5 to 10 g of triallyl isocyanurate (0.02 to
0.04 mol), 1 to 3 g of polymerization initiator such as
azobisisobutyronitrile are mixed, and the resultant organic solvent
is suspended in 500 to 1000 mL of 0.01 to 1.0% by weight of methyl
cellulose aqueous solution, and is reacted at 60 to 80.degree. C.
for 5 to 10 hours. By this method, the prepolymer relevant to the
present invention can be obtained. After the reaction, the
resultant polymer relevant to the present invention is washed with
water or methanol if necessary. Subsequently, 10 g of prepolymer
relevant to the present invention is suspended in 50 to 100 mL of
20 to 50% by volume of 2-propanol aqueous solution, and 10 to 50 g
of N,N-dimethylaminoethane (0.1 to 0.5 mol) is added, and the
organic solution is reacted at 40 to 60.degree. C. for 10 to 20
hours while stirring. Then, the polymer of the present invention
can be obtained.
[0048] The acidic groups of the perfluoro compound having the
acidic group relevant to the present invention include sulfo group,
carboxyl group, hydroxyl group and the like, and sulfo group,
carboxyl group are preferable. The perfluoro compounds having sulfo
group at terminal include the ones having 2 to 18 carbon atoms and
the like, preferably 4 to 10 and the like. The perfluoro compounds
having carboxyl group at terminal include the ones having 2 to 18
carbon atoms and the like, preferably 4 to 18, more preferably 4 to
12, further preferably 4 to 10 and the like. The perfluoro
compounds having hydroxy group at terminal include the ones having
2 to 18 carbon atoms and the like, preferably 4 to 10 and the like.
It should be noted that, the perfluoro compounds having the acidic
group relevant to the present invention also include the ones which
can form salts by substituting hydrogen ion of sulfo group or
carboxyl group at terminal of the above-described perfluoro
compounds having the acidic group with sodium ion, potassium ion or
ammonium ion. Preferable specific examples include, for example,
perfluoro sulfonic acids such as perfluoro butane sulfonic acid
(C.sub.4F.sub.9--SO.sub.3H, abbreviation: PFBS), perfluoro hexane
sulfonic acid (C.sub.6F.sub.13--SO.sub.3H, abbreviation: PFHxS),
perfluoro octane sulfonic acid (C.sub.8F.sub.17--SO.sub.3H,
abbreviation: PFOS), perfluoro decane sulfonic acid
(C.sub.10F.sub.21--SO.sub.3H, abbreviation: PFDS); perfluoro
carboxylic acids such as perfluoro butanoic acid
(C.sub.4F.sub.9--COOH, abbreviation: PFBA), perfluoro pentanoic
acid (C.sub.5F.sub.11--COOH, abbreviation: PFPeA), perfluoro
hexanoic acid (C.sub.6F.sub.13--COOH, abbreviation: PFHxA),
perfluoro heptanoic acid (C.sub.7F.sub.15--COOH, abbreviation:
PFHpA), perfluoro octanoic acid (C.sub.8F.sub.17--COOH,
abbreviation: PFOA), perfluoro nonanoic acid
(C.sub.9F.sub.19--COOH, abbreviation: PFNA), perfluoro decanoic
acid (C.sub.10F.sub.21--COOH, abbreviation: PFDA), perfluoro
undecanoic acid (C.sub.11F.sub.23--COOH, abbreviation: PFUnDA),
perfluoro dodecanoic acid (C.sub.12F.sub.25--COOH, abbreviation:
PFDoDA), perfluoro tetradecanoic acid (C.sub.14F.sub.29--COOH,
abbreviation: PFTeDA), perfluoro hexadecanoic acid
(C.sub.16F.sub.33--COOH, abbreviation: PFHexDA), perfluoro
octadecanoic acid (C.sub.18F.sub.37--COOH, abbreviation:
PFODA).
[0049] The filler for measuring the perfluoro compound having the
acidic group of the present invention is the one containing the
polymer of the present invention obtained as described above.
[0050] The preprocessing column for measuring the perfluoro
compound having the acidic group (hereinafter, this may be
abbreviated as the preprocessing column for PF of the present
invention) is the one filled with the above-described filler for
measuring the perfluoro compound having the acidic group of the
present invention. Filling method and the like is not particularly
limited, and, in the column container normally used in this field,
the above-described filler may be filled according to the known
method, for example, slurry method, dry type-tapping method and the
like. Amount of the filler may be determined according to the
amount of use of the sample, for example, when 1000 ml of samples
of environmental water such as river water, lake water, sea water;
tap water, well water, spring water, body fluid and the like are
passed through column to adsorb the perfluoro compound having the
acidic group, amount of the filler is normally 10 to 1000 mg,
preferably 50 to 250 mg. Column volume may be appropriately
selected according to the filling material.
[0051] The measuring method of the perfluoro compound having the
acidic group using the preprocessing column for PF of the present
invention may be carried out by the following steps:
[0052] (1) a step of passing the sample containing the perfluoro
compound having the acidic group through the preprocessing column
for PF of the present invention to adsorb the perfluoro compound
having the acidic group;
[0053] (2) a step of eluting the perfluoro compound having the
acidic group by eluate; and
[0054] (3) a step of measuring the obtained perfluoro compound
having the acidic group.
[0055] In addition, in case of the sample having much impurities
such as river water, lake water, the preprocessing column for PF of
the present invention may be washed after adsorbing the perfluoro
compound having the acidic group in order to remove these
impurities from the preprocessing column for PF of the present
invention.
[0056] Detail of the measuring method of the perfluoro compound
having the acidic group using the preprocessing column of the
present invention is specifically described as bellows. That is,
first,
[0057] (1) when the sample containing the perfluoro compound having
the acidic group is passed through the preprocessing column for PF
of the present invention to adsorb the perfluoro compound having
the acidic group onto the column (filler), method of passing the
liquid may be free fall, or may be carried out by pressurization or
depressurization, and, pressurization or depressurization method is
preferable. When pressurization or depressurization method is
employed, adsorption rates are decreased when flow rates of sample
become too high, and therefore, flow rates of sample are determined
normally at to 30 mL/min, preferably at 10 to 30 mL/min, more
preferably at 10 to 20 mL/min. In addition, it is considered that
the perfluoro compound having the acidic group is held by ion
exchange capacity of the anion exchanging group in the polymer used
as the filler of the present invention, and therefore, when the
sample has basicity such as pH 8 or more, it is preferable to pass
the liquid after adjusting pH to 7 or less by the known
neutralization reaction. When washing is carried out after
operation of (1), washing solution may be passed through column,
and said washing solutions include aqueous organic solvents having
pH 8 or less, such as methanol, ethanol, propanol, acetonitrile,
acetone. The amount of the solvent may be appropriately determined
according to the amount of the filler, and it is normally 0.5 to 10
mL, preferably 0.5 to 5 mL relative to 10 mg of filler. Adjustment
of pH is not particularly limited if it is carried out by using the
acidic compounds used normally in this field, and the acidic
compounds include, preferably, for example, hydrochloric acid,
nitric acid, sulfuric acid and the like. Method of passing the
liquid may be free fall, or may be carried out by pressurization or
depressurization, and, pressurization or depressurization method is
preferable because the washing solution can be removed perfectly.
Next,
[0058] (2) when the perfluoro compound having the acidic group
adsorbed onto the preprocessing column for PF of the present
invention are eluted by eluate, the eluates include a buffer
solution having pH 8 to pH 14, preferably 9 to 14, a basic aqueous
solution, an aqueous organic solvent such as methanol, ethanol,
propanol, acetonitrile, acetone, and a mixed aqueous solution of
said aqueous organic solution and basic aqueous solution or buffer
solution, and the like since it is considered that the perfluoro
compound having the acidic group is held by ion exchange capacity
of the anion exchanging group in the polymer used as the filler of
the present invention. Adjustment of pH is not particularly limited
as long as it is carried out by using the basic compounds normally
to be used in this field, and the basic compounds include,
preferably, for example, ammonia. When ammonia is used,
concentration of ammonia is normally 0.1 to 2 w/w % by weight,
preferably 0.1 to 1 w/w % as concentration in eluate. In addition,
amount of use of eluate may be appropriately determined according
to the amount of the filler, and may be normally 0.3 to 100 mL,
preferably 0.3 to 10 mL, more preferably 0.3 to 1 mL relative to 10
mg of the filler. Further,
[0059] (3) By measuring the eluate containing the eluted perfluoro
compound having the acidic group with the known separation
measurement method, amount or concentration of the perfluoro
compound having the acidic group in the sample can be obtained. If
necessary, after the eluate containing the perfluoro compound
having the acidic group which is used for separation measurement,
is condensed by the known method, the elute may be provided with
the separation measurement method. The separation measurement
methods include, for example, liquid chromatography method,
electrophoretic method and the like. UV detection, fluorescence
detection, MS, MS/MS, evaporative light scattering detection (ELSD)
and the like known in itself may be used as the detection tool, and
measurement condition and the like may be determined according to
the one which is normally employed in this field. Among said
separation measurement methods, LC/MS/MS is preferable considering
from the point of separation and sensibility.
[0060] More specifically, for example, the measuring method is
carried out as follows. That is, for example, when 1000 mL of the
sample having 0.1 ng to 1 mg/1 L, preferably 0.1 ng to 1 .mu.g/1 L,
more preferably 0.1 to 100 ng/1 L of perfluoro compound having the
acidic group is used, the column filled with normally 10 to 1000
mg, preferably 50 to 200 mg of the filler of the present invention
for measuring the perfluoro compound having the acidic group is
used, and the sample is passed through said column by pressuring or
depressuring at flow rates of 5 to 20 ml/min. Subsequently, the
column is washed by passing 5 to 10 mL of purified water having pH
6 to 8 at flow rates of 5 to 10 mL/min, using pressuring or
depressuring method. Next, by using 2 to 6 mL of methanol having
0.1 to 1% of ammonia, it is eluted by free fall. By measuring the
obtained eluate with LC/MS/MS, under the condition which is
normally employed in this field, the perfluoro compounds having the
acidic group in the sample can be measured.
[0061] The drugs relevant to the present invention include all the
materials having medicinal properties, and the doping substance
such as diuretic and the other concealment drug, stimulant drug,
narcotic, p-blocker, specific material listed in The 2008
Prohibited List International Standard (World Anti-Doping Code),
2008 Monitoring Program and the like, and the medicament or drug,
which can temporally heighten or lessen the racing performance of
race-horse, described in the Attached Table (2) in Ordinance for
Enforcement of Horse Racing of The Japan Racing Association of the
Japan Racing Association Rule No. 2 effective in Heisei 19 year
(2007). Among them, the medicament or drug, which can temporally
heighten or lessen the racing performance of race-horse, are
preferable, specifically, for example, Atoropine, Amobarbital,
Allobarbital, Antipyrine, Oxyethyltheophylline,
Oxypropyltheophylline, 3'-Oxohexobarbital, Caffeine,
Chlorpromazine, Cocaine, Cyclobarbital,
.beta.,.gamma.-Dihydroxydibucaine, Dibucaine, Cyproheptadine,
Cyproheptadine-N-oxide, Dimorphoramine, Scopolamine, Strychnine,
Secobarbital, Theophylline, Tetracaine, Nikethamide, Nicotine,
Noscapine, Barbital, 3'-Hydroxyamobarbital,
.gamma.-Hydroxydimorphoramine, 3'-Hydroxysecobarbital,
3'-Hydroxyhexobarbital, 3'-Hydroxypentobarbital, Pipradrol,
Phenacetin, Phenylethylmalondiamide, Phenylbutazone, Phenobarbital,
Primidone, Brucaine, Procaine, Promazine, Hexobarbital,
Pentazocine, Pentetrazol, Pentobarbital, Methapyrilene,
Methamphetamine, Metharbital, Methylephedrine, Methylphenidate,
Methoxyphenamine, Mephobarbital, Monoethylglycinexylidide,
Lidocaine, Chlorpromazine sulfoxide, Nortriptylin, Salicylic acid,
Naproxen, and the like. Among them, Theophylline, Scopolamine,
Caffeine, Atoropine, Barbital, Nortriptylin, Salicylic acid,
Naproxen and the like are preferable.
[0062] The filler for measuring the drug of the present invention
is the one containing the polymer of the present invention obtained
as described above.
[0063] The preprocessing column for measuring the drugs of the
present invention is the one filled with the above-described filler
for measuring the drug of the present invention. Filling method and
the like is not particularly limited, the above-described filler
may be filled into the column container conventionally used in this
field, according to the known method, for example, slurry method,
dry type-tapping method and the like. The amount of filler in the
column may be determined according to the amount of sample to be
used, for example, when 10 mL of the sample having 0.001 to 50
.mu.g/mL, preferably 0.05 to 10 .mu.g/mL of drug is passed through
column to adsorb the drug, the amount of the sample is normally 10
to 1000 mg, preferably 50 to 200 mg, more preferably 50 to 100 mg.
Column volume may be appropriately selected according to the amount
of filler.
[0064] Measuring method of the drugs using the preprocessing column
for measuring the drugs of the present invention may be carried out
by the following steps:
[0065] (1) a step of passing the sample containing the drugs
through the preprocessing column for measuring drugs of the present
invention to adsorb the drugs;
[0066] (2) a step of eluting the drugs by eluate; and
[0067] (3) a step of measuring the eluted drugs.
[0068] In addition, in case of the sample having much impurities,
treatment such as washing may be carried out after adsorbing the
drugs in order to remove these impurities from the preprocessing
column for measuring the drugs of the present invention.
[0069] Detail of the measuring method of the drug using the
preprocessing column for measuring the drug of the present
invention is specifically described as follows. That is, first,
after adjusting the conditioning of the column by the known
method,
[0070] (1) The sample containing the drug is passed through the
preprocessing column for measuring the drug of the present
invention to adsorb the drug onto the column (filler). Method of
passing the liquid may be free fall, or may be carried out by
pressurization or depressurization, and pressurization or
depressurization method is preferable. When pressurization or
depressurization method is employed, adsorption rate is decreased
when flow rates of the sample become too high. Therefore, flow
rates of the sample are determined normally at 1 to 30 mL/min,
preferably at 10 to 30 mL/min, more preferably at 10 to 20 mL/min.
It should be noted that, when pH of the sample having the acidic
drug is 8 or more, it is preferable to pass the liquid after
adjusting pH of the sample to 7 or less by the known neutralization
reaction. That is, it is considered that the acidic drugs are held
by ion exchange capacity of the anion exchanging group in the
polymer of the present invention, and therefore, in order to
prevent the reduction of adsorption rate, pH of the sample is
preferably adjusted to 7 or less. When washing is carried out after
operation of (1), washing solution may be passed through column. As
the washing solution, buffer solution having of pH 6 to 8 is used,
and said buffer solution may be the one having 5 to 10% of aqueous
organic solvent such as methanol, ethanol, propanol, acetonitrile,
acetone, dichloromethane. The liquid amount of washing solution may
be appropriately determined according to the amount of the filler,
and it is normally 0.5 to 10 mL, preferably 0.5 to 5 mL relative to
10 mg of filler. It should be noted that, pH adjustment of washing
solution may be carried out according to the method normally used
in this field. Method of passing the liquid may be free fall, or
may be carried out by pressurization or depressurization, and
passing the liquid by pressurization or depressurization is
preferable because the washing solution can be removed
perfectly.
[0071] Next,
[0072] (2) when the drugs adsorbed onto column are eluted by
eluate, the neutral and basic drugs are eluted by the aqueous
organic solvent such as methanol, ethanol, propanol, acetonitrile,
acetone, dichloromethane, and acidic drug is eluted by the acidic
aqueous solution or the acidified aqueous organic solvent such as
methanol, ethanol, propanol, acetonitrile, acetone, dichloromethane
having normally pH 1 to 3, preferably pH 1 to 2, and the like. pH
adjustment of said acidic aqueous solution or acidic aqueous
organic solvent is not particularly limited, as long as it is
carried out by the method using the acidic compound normally used
in this field. Said acidic compounds include, preferably, for
example, hydrochloric acid, nitric acid, acetic acid, formic acid
and the like. A specific adjusting method is carried out as
follows. For example, when pH is adjusted to 1 to 2 by using formic
acid, the acidic aqueous solution or the acidic aqueous organic
solvent may be adjusted so that formic acid may contain normally 1
to 5 w/w %, preferably 1 to 2 w/w % as a concentration in the
eluate. Amount of the eluate in the elution of the above-described
(2) may be appropriately determined according to the amount of
filler, and it is normally 0.3 to 100 mL, preferably 0.3 to 10 mL,
more preferably 0.3 to 1 mL relative to 10 mg of the filler.
[0073] Further,
[0074] (3) by measuring the eluate containing the eluted drugs with
the known separation measurement method, amount or concentration of
the drug in the sample can be obtained. If necessary, after the
eluate containing the drug, which is used in separation
measurement, is condensed by the known method, the elute may be
provided with the separation measurement method. The separation
measurement methods include, for example, liquid chromatography
method, gas chromatography method, electrophoretic method and the
like, and as detection tool, UV detection, fluorescence detection,
MS, MS/MS, evaporative light scattering detection (ELSD) and the
like known in itself may be used, and measurement condition and the
like may be determined according to the one which is normally
employed in this field.
[0075] More specifically, for example, the measurement method is
carried out as follows. That is, for example, when 10 mL of the
sample having 1 to 10 .mu.g/mL of drugs is used, the column filled
with 50 to 100 mg of the filler for measuring the drug of the
present invention is used. 2 to 5 mL of aqueous organic solvent
such as methanol, 2 to 5 ml of purified water, and 2 to 5 mL of
buffer solution of sodium acetate having pH 6 to 8 are sequentially
passed through said column, and thus, column condition is adjusted.
Next, 10 mL of the sample is passed through the column. Then, it is
washed by passing 1 to 3 mL of the buffer solution of 0.1 to 1
mol/L of sodium acetate having 5 to 10% of methanol. Next, 2 to 5
mL of methanol is passed to elute the neutral and basic target
drug. Further, 2 to 5 mL of methanol having pH 1 to 2 is passed
through to elute the acidic target drugs. Each of the obtained
eluates is separated with HPLC at the setting condition which is
normally carried out in this field, and the target drugs are
measured by UV, fluorescent detector and the like, and therefore,
the amount of the target drug in the sample is obtained. It should
be noted that, the acidic drugs, and the neutral and basic target
drugs may be eluted into one container, and the acidic, neutral and
basic drugs may be simultaneously analyzed by separation.
[0076] Next, the present invention is described in more detail by
the Examples. However the present invention is not limited by these
Examples.
EXAMPLE 1
Synthesis of Polymer-1 of the Present Invention
[0077] As a hydrophobic monomer, 85.0 g of divinylbenzene (produced
by Nippon Steel Chemical Co., Ltd.) (purity:80%) (containing 0.52
mol of pure divinylbenzene), as a polar monomer (monomer in which
ion exchanging group can be introduced), 15.0 g (0.106 mol) of
glycidyl methacrylate (produced by Wako Pure Chemical Industries,
Ltd) were used, and as a water-insoluble organic solvent, 100 g of
toluene was used. 1.0 g of azobisisobutyronitrile (produced by Wako
Pure Chemical Industries, Ltd.) was added to mixed solvent of 2
kinds of the above-described monomers and the water-insoluble
organic solvent, and dissolved. Then, mixed solution was suspended
in 0.8 L of 0.2% by weight aqueous solution of methylcellulose, and
was stirred at high speed to adjust the average diameter of oil
droplet to about 60 .mu.m. Then, mixed solution was transferred to
polymerization apparatus equipped with stirrer, and was reacted at
80.degree. C. for 6 hours to obtain the cross-linked porous
copolymerized particles. These particles were washed with 2 L of
water and 2 L of methanol after filtering, and were dipped in ethyl
acetate overnight. Then, these particles were washed with 2 L of
methanol and dried. The resultant cross-linked porous copolymerized
particles had 10.26 nm of average micropore diameter and 636
m.sup.2/g of specific surface area. These particles were classified
with 32 .mu.m and 63 .mu.m sieves.
[0078] 10.0 g of porous copolymer after classification was
dispersed in 50 mL of 50% aqueous solution of 2-propanol to provide
homogeneous slurry, and was transferred to the reaction apparatus
equipped with stirrer, and amination reaction was carried out by
adding 18.5 g (0.21 mol) of morpholine (produced by Wako Pure
Chemical Industries, Ltd) at 50.degree. C. for 20 hours. After
reaction completed, reaction product was sequentially washed with
water and methanol, and was dried, thus, the resultant polymer was
defined as polymer-1 of the present invention. It should be noted
that, content of morpholino group in the resultant polymer-1 of the
present invention was 0.45 mmol/g. In addition, structure of
polymer-1 of the present invention is shown as follows.
##STR00027##
EXAMPLE 2
Synthesis of Polymer-2 of the Present Invention
[0079] Cross-linked porous copolymerized particles were obtained
using the similar procedure to Example 1. The obtained cross-linked
porous copolymerized particles had 10.56 nm of average micropore
diameter and 662.5 m.sup.2/g of specific surface area. These
particles were classified using 32 .mu.m and 63 .mu.m sieves.
Diethylamino group was introduced to the porous copolymer by the
similar procedure to Example 1 except that 15.5 g of diethylamine
(produced by Wako Pure Chemical Industries, Ltd) was used instead
of 18.5 g of morpholine, and thus, the resultant polymer was
defined as polymer-2 of the present invention. It should be noted
that, content of diethylamino group in the obtained polymer-2 of
the present invention was 0.56 mmol/g. In addition, structure of
polymer-2 of the present invention is shown as follows.
##STR00028##
EXAMPLE 3
[0080] Addition and Recovery Experiment of the Perfluoro Compound
having the Acidic Group (PFCs) Using Polymer-1 of the Present
Invention, Polymer-2 of the Present Invention
1. Preparation of Column and Sample
1-1. Preparation of Column
[0081] Each of 60 mg of Polymer-1 and 60 mg of polymer-2 of the
present invention prepared in Example 1 and Example 2 was filled
into the 3 ml of polypropylene syringes (manufactured by Pronics
Co. Ltd) equipped with filter having 20 .mu.m of micropore diameter
(Yoko Co., Ltd) at upper and lower ends respectively. These were
defined as solid-phase extraction column-1 and solid-phase
extraction column-2.
1-2. Preparation of the Sample
[0082] In 1000 ml of the purified water, 13 kinds of the perfluoro
compounds having the acidic group [perfluoro sulfonic acids: PFBS
(produced by Wako Pure Chemical Industries, Ltd); PFHx (produced by
Wellington Co., Ltd); PFOS (produced by Accustandard, Inc.); PFDS
(produced by Wellington Co., Ltd); perfluoro carboxylic acid:
PFPeA, PFHxA, PFHpA, PFOA, PFNA, PFDA, PFUnDA (produced by Wako
Pure Chemical Industries, Ltd), PFDoDA (produced by Hydrus Chemical
Inc), PFTeDA (produced by Fluorochem Ltd)] were added by 20 ng
respectively, and each was defined as sample-water.
[0083] As the above-described purified water, the purified water
which was passed through the Presep-C PFC (Short: produced by Wako
Pure Chemical Industries, Ltd) subjected to conditioning was used.
It should be noted that, the purified water in the Examples of this
application was the one treated by the same procedure.
[0084] In addition, each of 13 kinds of the perfluoro compounds
having the acidic group (perfluorosulfonic acids: PFBS, PFHxS,
PFOS, PFDS; perfluoro carboxylic acids: PFPeA, PFHxA, PFHpA, PFOA,
PFNA, PFDA, PFUnDA, PFDoDA, PFTeDA) was dissolved in methanol to
prepare the solution having by 20 ng/2 mL respectively. Each of
methanolic solutions containing the obtained 13 kinds of the
perfluoro compound having the acidic group respectively was defined
as standard sample.
2. Solid-Phase Extraction by Column
(1) Conditioning of the Column
[0085] Conditioning was carried out by passing 5 ml of methanolic
solution having 0.1% of ammonia, 5 mL of methanol (methanol for
LC/MS: produced by Wako Pure Chemical Industries, Ltd), and 10 ml
of purified water sequentially by free fall respectively into
solid-phase extraction column-1 and solid-phase extraction
column-2.
(2) Passing the Sample Solution
[0086] 1000 mL of the sample water was passed through by using
aspiration manifold (manufactured by JT Baker Inc) in which
depressurization force was set so as to obtain 10 to 20 mL/min of
flow rates.
(3) Washing
[0087] By using aspiration manifold (manufactured by JT Baker Inc)
in which depressurization force was set so as to obtain 1 to 2
mL/min of flow rates, solid-phase extraction column was washed with
5 mL of purified water, and further, water was removed from the
column while aspirating for 5 minutes.
(4) Elution
[0088] Two mL of the methanolic solution having 0.1% ammonia was
passed by 2 times free fall and eluted, and each of eluates
obtained by 2 times elution was taken into 2 mL of graduated
disposable spits (manufactured by Falcon Co. Ltd) made of
polypropylene (made of PP) respectively.
(5) Preparation of the Test Solution
[0089] To each of the obtained eluates, methanol (methanol for
LC/MS: produced by Wako Pure Chemical Industries, Ltd) was added to
adjust to 2 mL of volume, and this solution was defined as the test
solution. In addition, all the used apparatuses were reused after
washing with methanol and drying in order to remove contamination
of PFCs
3. Measurement Results by LC/MS/MS Method
[0090] Two test solutions (first eluate and second eluate) obtained
from the above-described operation of solid-phase extraction and
the standard samples were analyzed by LC/MS/MS. Area value obtained
from analysis of the standard samples was specified as 100%, and
recovery rate of the test solutions was determined. These results
were shown in Table-1. In addition, measurement condition of
LC/MS/MS was shown as follows:
[HPLC Condition]
[0091] Column: Wakopak Navi C18-5, 2.0.times.150 mm [0092]
(manufactured by Wako Pure Chemical Industries, Ltd);
[0093] Eluate: A) 10 mM ammonium acetate aqueous solution; [0094]
B) acetonitrile;
[0095] Time program: 0-25 min B=35-90%, 25-30 min B=90%, [0096]
30-35 min B=90-35%, 35-40 min B=35%;
[0097] flow rates: 0.2 mL/min, column temperature: 40.degree.
C.;
[0098] amount of injection: 5 .mu.L;
[LC/MS/MS: Ionization Condition]
[0099] Curtain Gas (CUR): 10;
[0100] Collision Gas (CAD): 5;
[0101] Ion Spray Voltage (IS): -4500;
[0102] Temperature (TEM): 400;
[0103] Ion Source Gas1 (Gas1): 80;
[0104] Ion Source Gas2 (Gas2): 70;
[0105] Each detective ion of the perfluoro compounds having the
acidic group (Q1 and Q3) is shown as follows:
TABLE-US-00001 TABLE 1 Recovery rate (%) by Recovery rate (%) by
LC/MS/MS solid-phase solid-phase Detected extraction column-1
extraction column-2 Ion First Second First Second Compound (atomic
mass unite) elution elution Sum elution elution Sum 1 PFBS
298.8/79.6 96 2.2 99 95 0.8 96 amu 2 PFHxS 398.8/79.6 92 1.1 93 106
0.3 106 amu 3 PFOS 498.8/79.6 99 1.7 101 98 1.4 100 amu 4 PFDS
598.9/79.9 91 0.2 91 90 0.0 90 amu 5 PFPeA 262.8/219.1 102 1.6 104
98 0.8 99 amu 6 PFHxA 312.9/268.6 93 3.5 97 86 1.3 87 amu 7 PFHpA
362.8/318.7 98 3.3 101 97 2.6 99 amu 8 PFOA 412.9/368.9 102 5.8 108
100 4.3 104 amu 9 PFNA 462.7/418.8 102 0.8 102 102 0.0 102 amu 10
PFDA 512.9/469.0 99 0.9 100 83 0.0 83 amu 11 PFUnDA 562.9/519.0 --
-- -- 86 1.1 87 amu 12 PFDoDA 612.9/568.9 -- -- -- 81 0.2 81 amu 13
PFTeDA 712.9/669 -- -- -- 96 0.5 96 amu
[0106] As apparent from the above-described Table-1, as a result of
solid phase extraction using column-1 and 2 of the present
invention, it was found that various perfluoro compounds having the
acidic group can be extracted with 80% or more of high recovery
rate by using any columns. Further, in the column-1 of the present
invention, all of the 10 kinds of the perfluoro compounds having
the acidic group have shown 90% or more of recovery rate. Also, in
the column-2 of the present invention, the perfluoro sulfonic acids
have shown 90% or more of recovery rate. That is, even in any
columns, significantly high recovery rate was found to show in the
perfluoro compound having the acidic group having 10 or less of
carbon atoms. As for elution, it was found that target substances
were almost completely eluted by 2 mL of the first eluate in case
of 60 mg of the filler of the present invention because elution
effect by second elution (2 mL) was weak.
EXAMPLE 4
[0107] Addition and Recovery Experiment of the Perfluoro Compound
having the Acidic Group (PFCs) Using Polymer-1 of the Present
Invention
1. Preparation of Column and Sample
1-1. Preparation of Column
[0108] Into the 3 ml of polypropylene syringe (manufactured by
Pronics Co. Ltd) equipped with filter having 20 .mu.m of micropore
diameter (Yoko Co., Ltd), 60 mg of Polymer-1 of the present
invention prepared in Example 1 was filled. This was defined as
solid-phase extraction column-3
2. Solid-Phase Extraction by Column
[0109] Solid-phase extraction was carried out by the similar way to
Example 3 except that elution was one time.
3. Measurement Results by LC/MS/MS Method
[0110] The liquid obtained by the solid-phase extraction operation
and the standard samples were analyzed relatively by LC/MS/MS. Area
value obtained from analysis of the standard samples was specified
as 100%, and recovery rate of the test solution was determined.
These results were shown in Table-2. In addition, measurement
condition of LC/MS/MS was the same as Example 3.
TABLE-US-00002 TABLE 2 Recovery rate (%) by solid-phase extraction
Perfluoro compound column-3 1 PFBS 100 2 PFHxS 90 3 PFOS 92 4 PFDS
90 5 PFPeA 86 6 PFHxA 90 7 PFHpA 90 8 PFOA 92 9 PFNA 100 10 PFDA 92
11 PFUnDA 94 12 PFDoDA 92 13 PFTeDa 92
[0111] From the result of Table-2, it was found that only the
recovery rate of PFPeA (perfluoro pentanoic acid) was 86%, and all
of the other recovery rates were more than 90%, and all of the
perfluoro compounds having the acidic group can be recovered by 80%
or more when the polymer-1 of the present invention was employed.
Therefore, most of the compounds have shown 90% or more of recovery
rate.
COMPARATIVE EXAMPLE 1
Addition and Recovery Experiment of PFCs Using Presep-C Agri
(Short) Column
1. Preparation of Column and Sample
1-1. Column
[0112] Commercially available Presep-C Agri (Short) (220 mg, closed
type column, manufactured by Wako Pure Chemical Industries, Ltd)
was used. It should be noted that, structure of Presep-C Agri was
shown as follows:
##STR00029##
1-2. Preparation of Sample
[0113] To 1000 ml of the purified water, 13 kinds of the perfluoro
compounds having the acidic group (perfluoro sulfonic acid: PFBS,
PFHx, PFOS, PFDS, perfluoro carboxylic acid having the acidic
group: PFPeA, PFHxA, PFHpA, PFOA, PFNA, PFDA, PFUnDA, PFDoDA,
PFTeDA) were added by 20 ng respectively. This solution was defined
as sample water. Also, each of 13 kinds of the perfluoro compounds
having the acidic group (perfluorosulfonic acid: PFBS, PFHx, PFOS,
PFDS, perfluorocarboxylic acid: PFPeA, PFHxA, PFHpA, PFOA, PFNA,
PFDA, PFUnDA, PFDoDA, PFTeDA) was dissolved in methanol to adjust
the solution to 20 ng/2 mL respectively. Each solution was defined
as the standard sample.
2. Solid-Phase Extraction by Column
(1) Conditioning of Column
[0114] 10 mL of methanol (methanol for LC/MS, produced by Wako Pure
Chemical Industries, Ltd) and 10 mL of purified water were passed
sequentially through Presep-C Agri by free fall.
(2) Passing of the Sample Solution
[0115] 1000 mL of the sample water was passed through by using
aspiration manifold (manufactured by JT Baker Inc) in which
depressurization force was set so as to obtain 10 to 20 mL/min of
flow rates.
(3) Washing
[0116] By using aspiration manifold (manufactured by JT Baker Inc)
in which depressurization force was set so as to obtain 1 to 2
mL/min of flow rates, column was washed with 5 mL of purified
water, and further, aspirating was continued for 5 minutes to
remove water from the column.
(4) Elution
[0117] 2 mL of methanol (methanol for LC/MS: produced by Wako Pure
Chemical Industries, Ltd) was passed by free fall 2 times and
eluted, and each of eluates obtained by 2 times elution was taken
into 2 mL of graduated disposable spits made of PP
respectively.
(5) Preparation of the Test Solution
[0118] To each of the obtained eluates, methanol (methanol for
LC/MS: produced by Wako Pure Chemical Industries, Ltd) was added to
adjust 2 mL of volume respectively, and this solution was defined
as the test solution. In addition, all the used apparatuses were
reused after washing with methanol and drying in order to remove
contamination of PFCs
3. Measurement Results by LC/MS/MS Method
[0119] Two test solutions obtained from the above-described
operation of solid-phase extraction (first eluate and second
eluate) and standard samples were analyzed by LC/MS/MS. Area value
obtained from analysis of standard samples was specified as 100%,
and recovery rate of the test solutions were determined. These
results were shown in Table-3. In addition, measurement condition
of LC/MS/MS was the same as Example-3.
TABLE-US-00003 TABLE 3 Recovery rate by Presep-C Agri (Short) (%)
Perfluoro First Second Compounds Elution Elution Sum 1 PFBS 16 0.1
16 2 PFHxS 82 1.9 84 3 PFOS 93 0.2 93 4 PFDS 93 1.0 94 5 PFPeA 9
0.2 10 6 PFHxA 18 0.7 19 7 PFHpA 47 0.8 48 8 PFOA 92 1.4 93 9 PFNA
98 1.0 99 10 PFDA 93 1.1 94 11 PFUnDA 91 1.2 92 12 PFDoDA 85 1.4 87
13 PFTeDa 74 2.0 76
[0120] As apparent from results of the above-described Table-3, in
case of use of Presep-C Agri column containing the hydrophilic
reversed-phase polymer resin, each of recovery rates for perfluoro
sulfonic acids having 4 or less of carbon atoms, and perfluoro
carboxylic acids having 7 or less of carbon atoms was 50% or less.
In addition, recovery rate of PFTeDA having 14 carbon atoms was
less than 80%. Therefore, it was shown that Presep-C Agri column
can be used to solid-phase extraction of specified perfluoro
compounds having the acidic group, but it was unsuitable for
measuring simultaneously all the perfluoro compounds having the
acidic group.
COMPARATIVE EXAMPLE 2
[0121] Addition and Recovery Experiment of the Perfluoro Compound
having the Acidic Group (PFCs) Using Oasis HLB
1. Preparation of Column and Sample
1-1. Column
[0122] Commercially available Oasis HLB (225 mg, closed type:
manufactured by Waters Corp) was used.
1-2. Preparation of Sample
[0123] To 1000 ml of the purified water, 13 kinds of the perfluoro
compounds having the acidic group (perfluoro sulfonic acid: PFBS,
PFHx, PFOS, PFDS, perfluoro carboxylic acid: PFPeA, PFHxA, PFHpA,
PFOA, PFNA, PFDA, PFUnDA, PFDoDA, PFTeDA) were added by 20 ng
respectively. This solution was defined as sample water.
[0124] In addition, each of 13 kinds of the perfluoro compounds
having the acidic group (perfluorosulfonic acid: PFBS, PFHx, PFOS,
PFDS, perfluorocarboxylic acid: PFPeA, PFHxA, PFHpA, PFOA, PFNA,
PFDA, PFUnDA, PFDoDA, PFTeDA) was dissolved in methanol to adjust
the solution to 20 ng/4 mL respectively. Each solution was defined
as the standard sample.
2. Solid-Phase Extraction by Column
(1) Conditioning of Column
[0125] 5 ml of methanol (methanol for LC/MS: produced by Wako Pure
Chemical Industries, Ltd) and 5 ml of purified water were passed
sequentially by free fall into Oasis HLB. It should be noted that,
structure of filler of Oasis HLB was shown as follows.
##STR00030##
(2) Passing the Sample Solution
[0126] 1000 mL of the sample water was passed through by using
aspiration manifold (manufactured by JT Baker Inc) in which
depressurization force was set so as to obtain 10 to 20 mL/min of
flow rates.
(3) Washing
[0127] By using aspiration manifold (manufactured by JT Baker Inc)
in which depressurization force was set so as to obtain 1 to 2
mL/min of flow rates, column was washed with 5 mL of purified
water, and further, water was removed from the column while further
aspirating for 5 minutes.
(4) Elution
[0128] Four mL of methanol was passed by free fall and was eluted,
and eluate was taken into 4 mL of graduated disposable spits made
of PP.
(5)
[0129] To the obtained eluate, methanol (methanol for LC/MS:
produced by Wako Pure Chemical Industries, Ltd) was added to adjust
4 mL of volume. This solution was defined as test solution. In
addition, all the used apparatuses were reused after washing with
methanol and drying in order to remove contamination of PFCs
3. Measurement Results by LC/MS/MS Method
[0130] The test solution obtained by the above-described operation
of solid-phase extraction and the standard samples were analyzed by
LC/MS/MS respectively. Area value obtained from analysis of the
standard sample was specified as 100%, and recovery rates of the
test solution were determined. These results were shown in Table-4.
In addition, measurement condition of LC/MS/MS was the same as
Example 3.
COMPARATIVE EXAMPLE 3
[0131] Addition and Recovery Experiment of the Perfluoro Compound
having the Acidic Group (PFCs) Using Oasis WAX
1. Preparation of Column and Sample
1-1. Column
[0132] Commercially available Oasis WAX (225 mg, closed type,
manufactured by Waters Corp) was used.
1-2 Preparation of the Sample
[0133] To 1000 ml of the purified water, 13 kinds of the perfluoro
compounds having the acidic group (perfluoro sulfonic acid: PFBS,
PFHx, PFOS, PFDS, perfluoro carboxylic acid: PFPeA, PFHxA, PFHpA,
PFOA, PFNA, PFDA, PFUnDA, PFDoDA, PFTeDA) were added by 20 ng
respectively. Each solution was defined as sample water. In
addition, each of 13 kinds of the perfluoro compounds having the
acidic group (perfluorosulfonic acid: PFBS, PFHx, PFOS, PFDS,
perfluorocarboxylic acid: PFPeA, PFHxA, PFHpA, PFOA, PFNA, PFDA,
PFUnDA, PFDoDA, PFTeDA) was dissolved in methanol to adjust the
solution to 20 ng/4 mL respectively. Each solution was defined as
standard sample.
2. Solid-Phase Extraction by Column
(1) Conditioning of Column
[0134] Five ml of methanol (methanol for LC/MS: produced by Wako
Pure Chemical Industries, Ltd) and 5 ml of purified water were
passed sequentially by free fall into Oasis WAX. It should be noted
that, structure of filler of Oasis WAX was shown as follows.
##STR00031##
(2) Passing the Sample Solution
[0135] By using aspiration manifold (manufactured by JT Baker Inc)
in which depressurization force was set so as to obtain 10 to 20
mL/min of flow rates, 1000 mL of the sample water was passed
through.
(3) Washing
[0136] By using aspiration manifold (manufactured by JT Baker Inc)
in which depressurization force was set so as to obtain 1 to 2
mL/min of flow rates, column was washed with 2 mL of methanol, and
further aspirating was continued for 5 minutes to remove water from
column.
(4) Elution
[0137] Four ml of methanol having 1% of ammonia was passed through
by free fall and eluted, and eluate was taken into 4 mL of
graduated disposable spits (manufactured by Falcon Co. Ltd) made of
PP.
(5) Preparation of the Test Solution
[0138] To the obtained eluates, methanol (methanol for LC/MS:
produced by Wako Pure Chemical Industries, Ltd) was added to adjust
to 4 mL of volume, and this solution was defined as the test
solution. It should be noted that, all the used apparatuses were
used after washing with methanol and drying in order to remove
contamination of PFCs
3. Measurement Results by LC/MS/MS Method
[0139] The test solution obtained by the above-described operation
of solid-phase extraction and standard samples were analyzed by
LC/MS/MS respectively. Area value obtained from analysis of the
standard sample was specified as 100%, and recovery rate of the
test solution was determined. These results were shown in Table-4
in addition. It should be noted that, measurement condition of
LC/MS/MS was the same as Example 3.
TABLE-US-00004 TABLE 4 Perfluoro Recovery rate by Recovery rate by
compound Oasis HLB (%) Oasis WAX (%) 1 PFBS 92 85 2 PFHxS 94 92 3
PFOS 92 101 4 PFDS 80 84 5 PFPeA 90 88 6 PFHxA 96 90 7 PFHpA 94 92
8 PFOA 96 94 9 PFNA 96 104 10 PFDA 96 96 11 PFUnDA 92 90 12 PFDoDA
72 76 13 PFTeDa 50 64
[0140] As apparent from results of Table-4, in case of using Oasis
HLB and Oasis WAX, recovery rates of PFDoDA (perfluorododecyl
carboxylic acid) and PFTeDA (perfluorotetradecanoic acid) which
have 12 and over carbon atoms were less than 80%, particularly
PFTeDA having 14 carbon atoms (perfluorotetradecanoic acid) showed
low recovery rate less than 70% in any case.
[0141] As described above, it was found that, from results of
Table-3, Table-4, in case of using Presep-C Agri column in which
hydrophilic reversed polymer resin was used for recovering the
perfluoro compounds having the acidic group, recovery rate of the
compound having small carbon numbers (7 or less) or large carbon
numbers (14 or more) becomes low. In case of using commercially
available Oasis HLB, Oasis WAX for recovering the perfluoro
compounds having the acidic group, recovery rate of the compound
having large carbon numbers (12 or more) becomes low. On the other
hand, as apparent from results of Table-1, and Table-2, when the
columns of the present invention were used, 80% or more of recovery
rates were shown in all the perfluoro compounds having the acidic
group regardless of carbon numbers. Depending on the kind of the
introduced anion exchanging group, most of the perfluoro compounds
having the acidic group can be recovered by 90% or more of high
recovery rate. Therefore, it was found that the perfluoro compounds
having acidic group at terminal such as perfluoro sulfonic acid,
perfluoro carboxylic acid could be efficiently and simultaneously
measured regardless of carbon numbers when the column of the
present invention was used.
EXAMPLE 5
Synthesis of Polymer-3 of the Present Invention
[0142] As a hydrophobic monomer, 85.0 g of divinylbenzene (produced
by Nippon Steel Chemical Co., Ltd: purity:57%) (contained 0.37 mol
of pure divinylbenzene), as a polar monomer (monomer in which ion
exchanging group can be introduced), 15.0 g of glycidyl
methacrylate (containing 0.106 mol of glycidyl methacrylate)
(produced by Wako Pure Chemical Industries, Ltd) were used, and 100
g of toluene was used as a water-insoluble organic solvent. After
1.0 g of azobisisobutyronitrile (produced by Wako Pure Chemical
Industries, Ltd.) was added to this mixture and dissolved. Mixed
solution was suspended in 0.8 L of aqueous solution having 0.2% by
weight of methylcellulose, and was stirred at high speed to adjust
the average diameter of oil droplet at about 60 .mu.m. Then, mixed
solution was transferred to polymerization apparatus equipped with
stirrer and reacted at 80.degree. C. for 6 hours to obtain the
cross-linked porous copolymerized particles. These particles were
filtered, and were further washed with 2 L of water and 2 L of
methanol, and were dipped in ethyl acetate overnight. Then, these
particles were washed with 2 L of methanol and dried. It should be
noted that, the resultant cross-linked porous copolymerized
particles had 4.0 nm of average micropore diameter and 429
m.sup.2/g of specific surface area. Said particles were classified
with 32 .mu.m and 63 .mu.m sieves to obtain 37 g of particles
having about 40 .mu.m of average particle diameter.
[0143] After classification, 10.0 g of the obtained porous
copolymer was dispersed in 50 mL of aqueous solution having 50% of
2-propanol to provide homogeneous slurry, and was transferred to
the reaction apparatus equipped with stirrer, and amination
reaction was carried out by adding 15 g (0.17 mol) of
N,N-dimethylaminoethane (produced by Wako Pure Chemical Industries,
Ltd) at 40.degree. C. for 20 hours. After the reaction completed,
reaction product was sequentially washed with water and methanol,
and dried, and thus, polymer-3 of the present invention was
obtained. Content of introduced ethyldimethyl ammonium group (anion
exchanging group) was determined by reverse titration method, and
then amount of this group was shown as 0.34 meq/g gel dry. It
should be noted that, structure of polymer-3 of the present
invention is shown as follows.
##STR00032##
EXAMPLE 6
Synthesis of Polymer-4 of the Present Invention
[0144] The cross-linked porous copolymeric particles were obtained
by using the reagent and the procedure similar to Example 5 except
that as a hydrophobic monomer, 80.0 g of divinylbenzene (produced
by Nippon Steel Chemical Co., Ltd, purity:57%) (contained 0.37 mol
of pure divinylbenzene), as a polar monomer (monomer in which ion
exchanging group can be introduced), 15.0 g of glycidyl
methacrylate (contained 0.106 mol of glycidyl methacrylate)
(produced by Wako Pure Chemical Industries, Ltd) and triallyl
isocyanurate (produced by Tokyo Chemical Industry Co., Ltd)
(contained 0.02 mol of triallyl isocyanurate) were used. The
cross-linked porous copolymeric particles were washed by the
similar way, then dried. These cross-linked porous copolymeric
particles had 3.9 nm of average micropore diameter and 380
m.sup.2/g of specific surface area. After these particles were
classified by using 32 .mu.m and 63 .mu.m sieves,
ethyldimethylammonium group was introduced by the method similar to
Example-5, and polymer-4 of the present invention was obtained.
Content of the introduced ethyldimethylammonium group (anion
exchanging group) was determined by reverse titration method, and
amount of this group was 0.30 meq/g gel dry. It should be noted
that, structure of polymer-4 of the present invention was shown as
follows.
##STR00033##
COMPARATIVE EXAMPLE 4
Synthesis of Comparative Polymer-1
[0145] The cross-linked porous copolymeric particles were obtained
by using the reagent and the procedure similar to Example 5 except
that as a hydrophobic monomer, 65.0 g of divinylbenzene (produced
by Nippon Steel Chemical Co., Ltd, purity:57%) (contained 0.28 mol
of pure divinylbenzene), as a polar monomer (monomer in which ion
exchanging group can be introduced), 30.0 g of glycidyl
methacrylate (produced by Wako Pure Chemical Industries, Ltd)
(contained 0.21 mol of glycidyl methacrylate) and 5 g of
dimethylacrylamide (produced by Wako Pure Chemical Industries, Ltd)
(contained 0.05 mol of dimethylacrylamide), as a water-insoluble
organic solvent, 60 g of toluene, and 40 g of lauryl alcohol were
used. These resin particles were washed by the similar way, then
dried. These cross-linked porous copolymerized particles had 29.8
nm of average micropore diameter and 260 m.sup.2/g of specific
surface area. After classification of these particles with 32 .mu.m
and 63 .mu.m sieves, trimethylamino group was introduced by using
the reagent and the procedure similar to Example 5 except that
reaction time is 6 hours, and 40 g of trimethylamine (produced by
Wako Pure Chemical Industries, Ltd, 30% of aqueous solution)
instead of N,N-dimethylethaneamine was used, and comparative
polymer-1 was obtained. Amount of the introduced anion exchanging
group was determined by reverse titration method, and it was 0.29
meq/g gel dry. It should be noted that, structure of comparative
polymer-1 was shown as follows.
##STR00034##
COMPARATIVE EXAMPLE 5
Production of Comparative Polymer-2
[0146] As a hydrophobic monomer, 70.0 g of divinylbenzene (produced
by Nippon Steel Chemical Co., Ltd, purity:57%) (contained 0.306 mol
of pure divinylbenzene), as a polar monomer (monomer in which ion
exchanging group can be introduced), 20.0 g of chloromethylstyrene
(produced by Tokyo Chemical Industry Co., Ltd, purity:90%)
(contained 0.146 mol of pure chloromethylstyrene), as a polar
monomer, 10.0 g of dimethylacrylamide (produced by Wako Pure
Chemical Industries, Ltd) (contained 0.1 mol of dimethylacrylamide)
were used. As a water-insoluble organic solvent, 60 g of toluene,
40 g of lauryl alcohol were used. As polymerization initiator, 1.0
g of azobisisobutyronitrile (produced by Wako Pure Chemical
Industries, Ltd) was dissolved in this mixture, and then, mixed
solution was suspended in 0.8 L of aqueous solution having 0.2% by
weight of methylcellulose, and stirred at high speed to adjust the
average diameter of oil droplet to about 40 .mu.m. Then, mixed
solution was transferred to polymerization apparatus equipped with
stirrer, and reacted at 80.degree. C. for 6 hours to obtain the
cross-linked porous copolymerized particles. These particles were
filtered, and further washed with 2 L of water and 2 L of methanol,
and dipped in ethyl acetate overnight. Subsequently, these
particles were washed with 2 L of methanol, and dried. It should be
noted that, the cross-linked porous copolymeric particles obtained
by such method had 3.8 nm of average micropore diameter and 96
m.sup.2/g of specific surface area. Said particles were classified
by using 32 .mu.m and 63 .mu.m of sieves, and 35 g of particles
having about 40 .mu.m of average particle diameter were
obtained.
[0147] 10.0 g of the obtained porous copolymer after classification
was dispersed in 50 mL of aqueous solution having 50% of 2-propanol
to provide homogeneous slurry, and transferred to the reaction
apparatus equipped with stirrer, and amination reaction was carried
out by adding g of N,N-dimethylaminoethane (produced by Wako Pure
Chemical Industries, Ltd) at 40.degree. C. for 8 hours. After the
reaction completed, reaction product was sequentially washed with
water and methanol, and dried, and thus, comparative polymer-2 was
obtained. Amount of the introduced anion exchanging group was
determined by reverse titration method, and it was 0.34 meq/g gel
dry. It should be noted that, structure of comparative polymer-2
was shown as follows.
##STR00035##
EXAMPLE 7
Addition and Recovery Experiment of the Various Drug
[0148] To 6 mL of buffer solution of 2 mol/L of sodium acetate
adjusted at pH 7, all the 8 kinds of drugs shown as Table-5 were
added to prepare the test solutions containing the amount of drugs
shown in the table. It should be noted that, as for the drug poorly
soluble in the buffer solution of sodium acetate, once the drug of
amounts in the table were dissolved in methanol, then said methanol
was added to buffer solution.
[0149] In addition, into 2 mL of methanol, the amounts of 8 kinds
of the drugs described in the right column of in Table-1 were
added, and thus, the standard solution was prepared.
TABLE-US-00005 TABLE 5 Amount of Drug Amount of Drug in Test in
Standard Drug solution (.mu.g) Solution (.mu.g) Theophylline 5 5
Scopolamine 25 25 hydrobromide caffeine 5 5 Atropine sulfate 25 25
Barbital 10 10 Nortriptylin 10 10 Salicylic acid 20 10 Naproxen 20
10
[0150] As fillers, polymer-3, 4 of the present invention,
comparative polymer-1, 2, and Oasis MAX were used, and these
fillers were filled into the syringe type cartridges having 4 mL of
inner volume by 60 mg respectively. These cartridges were defined
as solid-phase extraction cartridges.
[0151] Addition and recovery experiments were carried out as
follows. That is, first, to the solid-phase extraction cartridge, 2
mL of methanol, 2 mL of purified water and 1 mL of buffer solution
having 2 mol/L of sodium acetate adjusted to pH 7 were passed, and
then, total volume of the above-described test solution was poured.
Subsequently, 2 mL of mixed solution of 95 mL of buffer solution
having 0.1 mol/1 L of sodium acetate adjusted to pH 7 and 5 mL of
methanol were passed to the same cartridge to wash. Then, 2 mL of
methanol was passed through solid-phase extraction cartridge to
elute the adsorbed materials adsorbed on solid-phase. 2 mL of
recovered solution was separated with HPLC, and by measuring UV
absorption thereof, and recovery rates were determined. The
obtained recovery rates (%) are shown in Table-2. It should be
noted that, value of UV absorption for which the standard solution
was directly separated and measured by HPLC at the same condition
was specified as standard value (100%), and the recovery rate was
calculated from the standard value (hereinafter, same). In
addition, HPLC condition was as follows.
[HPLC Condition]
[0152] Column: Wakopak Wakosil-II 5C18 HG, 4.6.times.150 mm;
[0153] Eluate: A) 0.1% of phosphoric acid B) acetonitrile;
[0154] Gradient: 0-15 min, B: 5-85%, 15-25 min: B: 5%;
[0155] Flow rates: 1.0 mL/min at 40.degree. C.;
[0156] Amount of injection: 5 .mu.l;
[0157] Apparatus: HPLC : Prominence system (Shimadzu);
[0158] Further, in order to elute the acidic compounds, 4 mL of
methanolic solution containing 2% of formic acid was passed
through. 4 mL of recovery solution eluted from the solid-phase
cartridge was separated by HPLC at the above-described condition,
and UV value thereof was measured, and then, recovery rates were
determined. The obtained recovery rates (%) are shown in
Table-3.
TABLE-US-00006 TABLE 6 Filler Addition Comp. Comp. Amount Poly-
Poly- Poly- Poly- Oasis (.mu.g/ mer-3 mer-4 mer-1 mer-2 MAX Drug
Column) Recovery rate (%) Neutral Theophylline 5 97.1 92.2 13.3
29.4 75.5 Com- caffeine 5 101.7 98.5 22.6 97.4 92.7 pound Barbital
10 102.7 99.1 38.0 86.4 93.5 Basic Scopolamine 25 106.6 103.0 50.7
58.7 94.0 com- hydrobromide pound Atropine 25 108.0 105.7 11.9 96.6
67.5 sulfate Nortriptylin 10 103.5 100.5 -- 101.0 87.0
hydrochloride Acidic Salicylic 20 0 0 -- 0 0 Com- acid pound
Naproxen 20 0 0 -- 0 0
TABLE-US-00007 TABLE 7 Filler Addition Comp. Comp. Amount Poly-
Poly- Poly- Poly- Oasis (.mu.g/ mer-3 mer-4 mer-1 mer-2 MAX Drug
Column) Recovery rate (%) Acidic Salicylic 20 106.8 104.6 -- 67.5
103.4 Com- acid pound Naproxen 20 83.9 86.9 -- 67.0 86.3
[0159] From the result of Table-6, it was found that by using
polymer-3 and polymer-4 of the present invention, even
Theophylline, Atropine sulfate, Nortriptylin hydrochloride and the
like which have shown low recovery rate when Oasis MAX was used,
can be recovered by almost 100% of recovery rate.
[0160] On the other hand, comparative polymer-1 having similar
structure in part to polymer-3 and 4 of the present invention has
shown as low recovery rate in any of neutral compound, basic
compound. Similarly, comparative polymer-2 having similar structure
in part to polymer-3 and 4 of the present invention has been shown
as low recovery rate in Theophylline, Scopolamine hydrobromide. It
is estimated that retention capability of neutral and basic
compounds is greatly affected depending on the structure of
polymer. Therefore, it was understood that the polymer structure
comprising divinylbenzene and glycidyl methacrylate and the polymer
structure comprising divinylbenzene, triallyl isocyanurate and
glycidyl methacrylate, like polymer-3 and polymer-4 of the present
invention, have contributed to high retention capability of neutral
and basic compound.
[0161] Though salicylic acid and Naproxen of acidic compound were
not eluted with methanol, as apparent from results of Table-7,
these compounds can be eluted by methanol containing 2% of formic
acid. According to these results, it was found that polymer-3 and 4
of the present invention have almost same recovery rate as Oasis
MAX. Comparative polymer-2 has less than 70% of recovery rate in
both salicylic acid and Naproxen, and it showed low value. Though
retention capability of acidic compound was estimated to depend
mainly on the anion exchanging group, according to this result, it
was found that even when the same ion exchanging group
(ethyldimethylammonium group) has been contained, retention
capability of acidic compound may vary depending on the different
structure of fundamental polymer structure. Therefore, it was found
that by introducing the anion exchanging group to the polymer like
polymer-3 and polymer-4 of the present invention, these polymers
have excellent retention capability for neutral and basic
compounds, as well as acidic compounds.
INDUSTRIAL APPLICABILITY
[0162] The present invention relates to provide the novel polymer,
and the filler comprising said polymer, and said filler can
efficiently adsorb the various perfluoro compounds having the
acidic group. Therefore, according to the solid-phase extraction
method using said fillers, various perfluoro compounds having the
acidic group can be simultaneously and efficiently measured. In
addition, fillers comprising the above-described polymer can
efficiently adsorb the drugs, and by solid-phase extraction method
using said fillers, acidic, basic or/and neutral compounds can be
efficiently measured.
* * * * *