U.S. patent number 3,880,590 [Application Number 05/481,913] was granted by the patent office on 1975-04-29 for test strip for ketone bodies.
This patent grant is currently assigned to Shionogi & Co., Ltd.. Invention is credited to Yasunao Ogawa, Ukio Yonetani.
United States Patent |
3,880,590 |
Ogawa , et al. |
April 29, 1975 |
Test strip for ketone bodies
Abstract
A test strip for the detection of ketone bodies, comprising an
absorbing material, a heavy metal salt and a nitroprusside salt.
The test strip is useful in the diagnosis of various diseases such
as diabetes, disorders in the digestive organs, renal
insufficiency, uremia, self-poisoning or malignant carcinoma.
Inventors: |
Ogawa; Yasunao (Ikeda,
JA), Yonetani; Ukio (Nara, JA) |
Assignee: |
Shionogi & Co., Ltd.
(Osaka, JA)
|
Family
ID: |
14915665 |
Appl.
No.: |
05/481,913 |
Filed: |
June 21, 1974 |
Foreign Application Priority Data
|
|
|
|
|
Nov 8, 1973 [JA] |
|
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48-125667 |
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Current U.S.
Class: |
422/420;
436/128 |
Current CPC
Class: |
G01N
33/64 (20130101); Y10T 436/200833 (20150115) |
Current International
Class: |
G01N
33/64 (20060101); G01n 031/22 (); G01n 033/16 ();
C09k 003/00 () |
Field of
Search: |
;23/253TP,23B
;252/408 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Wolk; Morris O.
Assistant Examiner: Turk; Arnold
Attorney, Agent or Firm: Wenderoth, Lind & Ponack
Claims
What we claim is:
1. A test strip for the detection of ketone bodies, comprising an
absorbing material, a heavy metal salt and a nitroprusside
salt.
2. A test strip for the detection of ketone bodies according to
claim 1, wherein the heavy metal salt is an inorganic salt of a
metal selected from the group consisting of iron, nickel, copper,
cobalt, manganese, chromium and zinc, and the nitroprusside salt is
a nitroprusside alkali metal salt.
3. A test strip for the detection of ketone bodies according to
claim 1, wherein the heavy metal salt is a member selected from the
group consisting of nickel chloride, ferric chloride, ferrous
chloride, ferric ammonium sulfate, ferrous sulfate, cuprous
chloride, cupric chloride, cobalt dichloride and cobaltic chloride,
and the nitroprusside salt is a member selected from the group
consisting of sodium nitroprusside and potassium nitroprusside.
4. A composition for the detection of ketone bodies, comprising a
heavy metal salt and a nitroprusside salt.
5. A composition for the detection of ketone bodies according to
claim 4, wherein the heavy metal salt is an inorganic salt of a
metal selected from the group consisting of iron, nickel, copper,
cobalt, manganese, chromium and zinc, and the nitroprusside salt is
a nitroprusside alkali metal salt.
6. A composition for the detection of ketone bodies according to
claim 4, wherein the heavy metal salt is a member selected from the
group consisting of nickel chloride, ferric chloride, ferrous
chloride, ferric ammonium sulfate, ferrous sulfate, cuprous
chloride, cupric chloride, cobalt dichloride, and cobaltic
chloride, and the nitroprusside salt is a member selected from the
group consisting of sodium nitroprusside and potassium
nitroprusside.
Description
This invention relates to a test strip for the detection of ketone
bodies and a composition therefor. More particularly, the present
invention relates to a test strip for the detection of ketone
bodies, comprising an absorbing material, a heavy metal salt and a
nitroprusside salt, and to a composition therefor comprising a
heavy metal salt and a nitroprusside salt.
It is known that ketosis takes place owing to insufficient intake
of sugars or disorder of sugar availability in the living bodies.
In patients with ketosis, ketone bodies such as acetoacetic acid,
acetic acid, acetone or .beta.-hydroxybutyric acid increase in the
blood, and they are excreted mainly as acetoacetic acid in the
urine. The ketosis indicates the presence of various diseases such
as diabetes, disorders in the digestive organs, renal
insufficiency, uremia, self-poisoning, malignant carcinoma, etc.
Thus, detection of ketone bodies in the body fluids such as urine,
serum or plasma can provide a diagnosis of such diseases.
Hitherto, a number of methods for the detection of ketone bodies
have been employed in the art. For example, a test strip comprising
a nitroprusside salt and an alkali substance is known for the
detection of ketone bodies. However, since nitroprusside salts are
very unstable to an alkali, such known method which is carried out
under alkaline conditions is not a good test method for the
detection of ketone bodies. Further, the test strips heretofore
employed in the art have drawbacks in that their production is
complicated and their sensitivity to ketone bodies is not high. As
a result of investigations to overcome such problems, the present
inventors have discovered that, without addition of a substance to
keep the system at an alkaline pH, use of a nitroprusside salt in
combination with a heavy metal salt provides easily a stable test
strip having a good sensitivity and specificity to ketone bodies,
particularly to acetoacetic acid. The present invention is based on
these discoveries.
The test strips and the compositions for the detection of ketone
bodies in accordance with the present invention can be prepared in
the following manner.
A nitroprusside salt and a heavy metal salt are dissolved or
suspended in a suitable solvent to give a solution. Then, an
absorbing material is dipped in the solution and dried in the air
or at a temperature of from room temperature to about 50.degree.C,
to give a test strip for the detection of ketone bodies. The
solution itself may be used in the detection of ketone bodies, but
a test strip prepared as above can be more conveniently used in
view of the preservation, stability and handling, rather than the
solution itself.
The heavy metal salt can be inorganic salts of a heavy metal of
which specific gravity is approximately more than 5. Examples of
the heavy metal are nickel, copper, cobalt, manganese, chromium and
zinc, and typical examples of such heavy metal salt are nickel
chloride, ferric chloride, ferrous chloride, ferric ammonium
sulfate, ferrous sulfate, cuprous chloride, cupric chloride, cobalt
dichloride and cobaltic chloride. Among them, the most preferred
heavy metal salt is nickel chloride. The metal salts can be used
usually at a concentration of 1-20%, preferably 5-15%.
As the nitroprusside salt, it is preferred to use a nitroprusside
inorganic salt such as alkali metal salts (e.g. sodium salt,
potassium salt). The nitroprusside salt can be usually used at a
concentration of 1-20%, especially 5-15%.
Examples of the solvent in preparing an impregnating solution
containing the heavy metal salt and the nitroprusside salt are
water, organic solvents (e.g. dimethylformamide, dimethylsulfoxide,
methanol, ethanol) and a mixture thereof.
The absorbing material to be dipped in a solution containing the
nitroprusside salt and the heavy metal salt can be a sheet of
paper, a piece of cloth and a stick of porous wood. Examples of the
kind of paper are filter paper, blotting paper, adsorbent paper
(e.g. silica gel paper, alumina paper), ion-exchange cellulose
paper (e.g. phosphonomethyl cellulose paper, sulfoethyl cellulose
paper, phospho cellulose paper, carboxymethyl cellulose paper,
guanidoethyl cellulose paper, diethylaminoethyl cellulose paper,
aminoethyl cellulose paper, "Ecteola"-cellulose paper which is an
epichlorhydrin, triethanolamine reacted cellulose ion exchange
paper, as note U.S. Pat. No. 3,598,704, p-aminobenzyl cellulose
paper, polyethyleneimine cellulose paper), ion-exchange Sephadex
paper and ion-exchange resin paper. The shape of the absorbing
material is not particularly limited, though it is usually used in
a strip form. Among such absorbing materials, ion-exchange
cellulose papers, particularly diethylaminoethyl cellulose paper,
can be preferably employed.
The solution itself containing the necessary ingredients may be
used to detect ketone bodies by adding it to a specimen of body
fluids such as urine, plasma or serum, whereby detectable color
change results. However, from the point of view of handling,
stability, and preservation, the composition is advantageously used
in the form of a solid prepareation, rather than the impregnating
solution itself. Solid preparations such as tablets, granules,
pills or powders can be prepared in a conventional manner. In the
production of such preparations, suitable additives (e.g.
excipient, disintegrating agent, dispersing agent, stabilizer,
binder) not affecting the color reaction may be added without
changing the basis of the present invention. The color change is
detectable only by dissolving such solid preparation in the body
fluids containing the ketone bodies. Also, the color reaction may
be effected in such a manner that a tablet containing a heavy metal
salt and a tablet containing a nitroprusside salt, normally
separately packed, are combined in a specimen of body fluids when
used. Usually, the test strip of the invention is used by dipping
it in a test sample or by impregnating a test sample into the test
strip, whereby detectable color change results. When plasma or
serum is used as a test sample, the same application is possible
using the test strip of the invention. The test strip of the
invention may be, if desired, used in a more convenient form such
as, for example, being held on a plastic sheet.
The test strip of the invention is sensitive to ketone bodies,
particularly to acetoacetic acid in the urine. Since characteristic
color reaction takes place depending on the concentration of the
ketone bodies to be detected, semi-quantitative detection for such
ketone bodies is possible. It is possible to detect acetoacetic
acid in an amount of more than 1 mg/100 ml in the urine with the
test strip of the invention. Other ketone bodies such as acetone or
.beta.-hydroxybutyric acid are generally not detectable with the
present test strip. In general, color tone of the test strip
depends on the kind of the heavy metal salts used. For example,
when nickel salts or cobalt salts; iron salts; and copper salts are
employed, the test strip turns purple; orange to brown; and blue to
purple, respectively, in the presence of acetoacetic acid. This
various color tones seem to be due to the formation of specific
colored chelate among the nitroprusside salt, acetoacetic acid and
heavy metal salt. It is considered that this chelate formation
contributes much to the stability of the nitroprusside salt, thus
giving a distinct and uniform color with strong intensity. When an
acidic urine sample is tested, it is preferred to use a test strip
containing additionally tris-(hydroxymethyl)-aminomethane having a
buffer action.
The following working examples will be given only to illustrate the
embodiments of the present invention, and it is to be construed
that the technical scope of the invention is not limited by the
examples given, many variations of which are possible without
departing from the concept of the present invention.
EXAMPLE 1
To a 5% aqueous solution (10 ml) of nickel chloride was added a 5%
dimethylformamide solution (3 ml) of sodium nitroprusside in water,
thus giving a turbid pale yellowish solution. A piece (5 .times. 6
cm) of diethylaminoethyl cellulose paper DE-81 (Whatman Co.) was
dipped in the solution for one minute and then the strip was dried
at room temperature in a dark place. Then, the strip was cut into
narrower pieces (1 .times. 0.8 cm) and the strips were each held on
a basic body such as a plastic sheet. The test strips thus obtained
were dipped independently in urine samples containing acetoacetic
acid of 1, 5, 10, 20, 50, and 100 mg/100 ml, respectively. The test
strip turned reddish purple with strong intensity within one
minute, depending on the concentration of acetoacetic acid of more
than 1 mg/100 ml. When the test strip was dipped in a test sample
wherein acetoacetic acid was not contained, the strip did not
change color and it retained its original color (pale yellow).
The test strip did not react with ascorbic acid or thiol compounds,
but it turned greenish blue in the presence of pyruvic acid and
pale brown in the presence of .alpha.-ketoglutaric acid.
EXAMPLE 2
A 5% dimethylformamide solution (3 ml) of sodium nitroprusside in
water was added to a 5% aqueous solution (10 ml) of ferric chloride
to give a pale yellowish solution. A piece (5 .times. 5 mc) of
silica gel paper M3F 8860 (Carl Chleicher & Chull Co.) was
dipped in this solution for one minute, dried in the air, cut into
pieces (1 .times. 1 cm) and held on a plastic sheet to give the
desired strips. The sensitivity of the test strip to acetoacetic
acid was the same with that of the test strip obtained in EXAMPLE
1.
EXAMPLE 3
Using a 10% aqueous ferrous chloride, ferric sulfate, copper
sulfate, cuprous chloride or cobalt dichloride in place of 5%
aqueous nickel chloride, the procedure of EXAMPLE 1 was followed to
give corresponding test strips.
The sensitivity of the test strips obtained above was the same with
that of the test strip obtained in EXAMPLE 1.
EXAMPLE 4
Using a filter paper (Toyo filter paper No. 131) or a
diethylaminoethyl cellulose paper SG-81 (Whatman Co.), the same
procedure as described in EXAMPLE 1 was carried out to give
corresponding test strips.
EXAMPLE 5
Using a 5% nickel chloride solution (10 ml) prepared with 0.3M
tris-(hydroxymethyl)-aminomethane solution (pH 6.8), the same
procedure as described in EXAMPLE 1 was carried out to give the
desired test strip.
* * * * *