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US3880590A - Test strip for ketone bodies - Google Patents

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Publication number
US3880590A
US3880590A US48191374A US3880590A US 3880590 A US3880590 A US 3880590A US 48191374 A US48191374 A US 48191374A US 3880590 A US3880590 A US 3880590A
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Prior art keywords
salt
test
nitroprusside
strip
metal
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Expired - Lifetime
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Yasunao Ogawa
Ukio Yonetani
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Shionogi and Co Ltd
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Shionogi and Co Ltd
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by the preceding groups
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/64Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving ketones
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00Chemistry: analytical and immunological testing
    • Y10T436/20Oxygen containing
    • Y10T436/200833Carbonyl, ether, aldehyde or ketone containing

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.

Description

United States Patent Ogawa et al.

1 Apr. 29, 1975 Foreign Application Priority Data Nov. 8, 1973 Japan 48-125667 U.S. Cl. 23/253 TP; 23/230 B; 252/408 Int. Cl.... G0ln 31/22; G0ln 33/16; CO9k 3/00 Field of Search 23/253 TP, 230 B; 252/408 References Cited UNITED STATES PATENTS 1/1940 Fortune 23/230 B 2,362,478 11/1944 Galat 23/230 B 2,509,140 5/1950 Free 23/230 B X 2,577,978 12/1951 Nicholls et a1. 23/230 B X 2,990,253 6/1961 Smeby 3,212,855 10/1965 Mast et a1. 3,598.704 8/1971 Dahlqvist 23/253 TP X Primary ExaminerMorris O. Wolk Assistant Examiner-Arnold Turk Attorney, Agent, or FirmWenderoth, Lind & Ponack 57 ABSTRACT v 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, selfpoisoning or malignant carcinoma.

6 Claims, N0 Drawings TEST STRIP FOR KETONE BODIES 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 B-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 50C, 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, euprous 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 1-207c, preferably 5-l57c.

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 l-207c, especially 5-157z.

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, paminobenzyl cellulose paper, polyethyleneimine cellulose paper), ion-exchange Sephadex paper and ionexchange 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 im-- pregnating 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/lOO ml in the urine with the test strip of the invention. Other ketone bodies such as acetone or B-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 aqueous solution 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 X 6 cm) of diethylaminoethyl cellulose paper DE-8l (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 l X 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 l, 5, 10, 20, 50, and 100 mg/lOO 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/lOO 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 a-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 X 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 X 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 I.

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 EXAM- PLE 1.

EXAMPLE 4 Using a filter paper (Toyo filter paper No. 131) or a diethylaminoethyl cellulose paper SG-8l (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 EX- AMPLE l was carried out to give the desired test strip.

What we claim is:

l. 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.

Claims (6)

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.
US3880590A 1973-11-08 1974-06-21 Test strip for ketone bodies Expired - Lifetime US3880590A (en)

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JP (1) JPS5143796B2 (en)
BE (1) BE816926A (en)
CA (1) CA1021237A (en)
DE (1) DE2432753C3 (en)
DK (1) DK142296C (en)
ES (1) ES427803A1 (en)
FR (1) FR2250999B1 (en)
GB (1) GB1451268A (en)
NL (1) NL162478C (en)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2390733A1 (en) * 1977-05-13 1978-12-08 Behringwerke Ag control solution for detection, to diagnosis, substances in the urine
US4147514A (en) * 1977-11-21 1979-04-03 Miles Laboratories, Inc. Test means and method for detecting ketone bodies
US4405721A (en) * 1980-03-22 1983-09-20 Behringwerke Aktiengesellschaft Diagnostic agent for the detection of ketone bodies
US4440724A (en) * 1982-11-01 1984-04-03 Helena Laboratories Corporation Composition for detecting ketone bodies and method of preparation
US4970172A (en) * 1986-12-22 1990-11-13 Abbott Laboratories Method and device for ketone measurements
US5071769A (en) * 1986-12-22 1991-12-10 Abbott Laboratories Method and device for ketone measurement
US5922283A (en) * 1997-07-07 1999-07-13 Teco Diagnostics Test strips for the determination of the ionic strength or specific gravity of an aqueous sample
US6149865A (en) * 1997-07-07 2000-11-21 Teco Diagnostics, Inc. Test strips for the determination of the ionic strength or specific gravity of an aqueous sample
US6541216B1 (en) 1999-12-22 2003-04-01 Roche Diagnostics Corporation Amperometric biosensor test strip
US6583722B2 (en) 2000-12-12 2003-06-24 Kimberly-Clark Worldwide, Inc. Wetness signaling device
US6603403B2 (en) 2000-12-12 2003-08-05 Kimberly-Clark Worldwide, Inc. Remote, wetness signaling system
US20100150778A1 (en) * 2005-06-23 2010-06-17 Siemens Healthcare Diagnostics Inc. Detection of Adulterated Samples

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4931404A (en) * 1986-12-22 1990-06-05 Abbott Laboratories Method and device for ketone measurement

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2186902A (en) * 1939-04-07 1940-01-09 Lilly Co Eli Urine acetone test
US2362478A (en) * 1941-12-24 1944-11-14 Denver Chemical Mfg Company Reagent for testing for acetone
US2509140A (en) * 1948-03-02 1950-05-23 Miles Lab Test reagent composition
US2577978A (en) * 1949-02-04 1951-12-11 Miles Lab Diagnostic composition
US2990253A (en) * 1959-05-21 1961-06-27 Miles Lab Diagnostic composition
US3212855A (en) * 1962-08-06 1965-10-19 Miles Lab Diagnostic device
US3598704A (en) * 1966-01-05 1971-08-10 Kabi Ab Diagnostic device for various sugars

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2186902A (en) * 1939-04-07 1940-01-09 Lilly Co Eli Urine acetone test
US2362478A (en) * 1941-12-24 1944-11-14 Denver Chemical Mfg Company Reagent for testing for acetone
US2509140A (en) * 1948-03-02 1950-05-23 Miles Lab Test reagent composition
US2577978A (en) * 1949-02-04 1951-12-11 Miles Lab Diagnostic composition
US2990253A (en) * 1959-05-21 1961-06-27 Miles Lab Diagnostic composition
US3212855A (en) * 1962-08-06 1965-10-19 Miles Lab Diagnostic device
US3598704A (en) * 1966-01-05 1971-08-10 Kabi Ab Diagnostic device for various sugars

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2390733A1 (en) * 1977-05-13 1978-12-08 Behringwerke Ag control solution for detection, to diagnosis, substances in the urine
US4172049A (en) * 1977-05-13 1979-10-23 Behringwerke Aktiengesellschaft Control-solution for diagnostic detection methods for substances contained in the urine
US4147514A (en) * 1977-11-21 1979-04-03 Miles Laboratories, Inc. Test means and method for detecting ketone bodies
DE2838675A1 (en) * 1977-11-21 1979-05-23 Miles Lab Test means and methods for evidence of ketone bodies
US4405721A (en) * 1980-03-22 1983-09-20 Behringwerke Aktiengesellschaft Diagnostic agent for the detection of ketone bodies
US4440724A (en) * 1982-11-01 1984-04-03 Helena Laboratories Corporation Composition for detecting ketone bodies and method of preparation
US4970172A (en) * 1986-12-22 1990-11-13 Abbott Laboratories Method and device for ketone measurements
US5071769A (en) * 1986-12-22 1991-12-10 Abbott Laboratories Method and device for ketone measurement
US5922283A (en) * 1997-07-07 1999-07-13 Teco Diagnostics Test strips for the determination of the ionic strength or specific gravity of an aqueous sample
US6149865A (en) * 1997-07-07 2000-11-21 Teco Diagnostics, Inc. Test strips for the determination of the ionic strength or specific gravity of an aqueous sample
US6541216B1 (en) 1999-12-22 2003-04-01 Roche Diagnostics Corporation Amperometric biosensor test strip
US6583722B2 (en) 2000-12-12 2003-06-24 Kimberly-Clark Worldwide, Inc. Wetness signaling device
US6603403B2 (en) 2000-12-12 2003-08-05 Kimberly-Clark Worldwide, Inc. Remote, wetness signaling system
US20100150778A1 (en) * 2005-06-23 2010-06-17 Siemens Healthcare Diagnostics Inc. Detection of Adulterated Samples
US8293188B2 (en) * 2005-06-23 2012-10-23 Siemens Healthcare Diagnostics, Inc. Detection of adulterated samples

Also Published As

Publication number Publication date Type
DE2432753B2 (en) 1981-01-22 application
NL7410509A (en) 1975-05-12 application
BE816926A (en) 1974-10-16 grant
CA1021237A (en) 1977-11-22 grant
JP869147C (en) grant
FR2250999A1 (en) 1975-06-06 application
NL162478C (en) 1980-05-16 grant
JPS5143796B2 (en) 1976-11-24 grant
BE816926A1 (en) grant
ES427803A1 (en) 1977-02-01 application
DK336474A (en) 1975-07-14 application
DK142296C (en) 1981-03-02 grant
DE2432753C3 (en) 1981-09-10 grant
DK142296B (en) 1980-10-06 grant
NL162478B (en) 1979-12-17 application
FR2250999B1 (en) 1977-10-07 grant
JPS5078394A (en) 1975-06-26 application
DE2432753A1 (en) 1975-05-15 application
CA1021237A1 (en) grant
GB1451268A (en) 1976-09-29 application

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