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Publication numberUS2290436 A
Publication typeGrant
Publication dateJul 21, 1942
Filing dateSep 26, 1940
Priority dateSep 26, 1940
Publication numberUS 2290436 A, US 2290436A, US-A-2290436, US2290436 A, US2290436A
InventorsKamlet Jonas
Original AssigneeMiles Lab
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Diagnostic composition and method
US 2290436 A
Abstract  available in
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Claims  available in
Description  (OCR text may contain errors)

Patented July 21, 1942 DIAGNOSTIC COMPOSITION AND .METHOD I Jonas Kamiet, Brooklyn, N. Y., assignor to Miles Laboratories, Inc., Eikhart, Ind., a corporation of Indiana No Drawing.

'lClaims.

The present invention relates to diagnostic compositions and, more particularly, to diagnostic compositions suitable for use in the qualitative detection and semi-quantitative estimation of blood in body fluids, such as urine, vomitus, gastric contents, semen, cerebrospinal fluid and in feces. It has for its object to provide a simple, rapid and convenient method for performing this test with a high degree of accuracy and specificity, and one that can readily be used by the average physician without laboratory equipment, supplies of fresh chemicals or specialized analytical training.

The detection of occult blood in body fluids and feces has become an invaluable aid to the medical practitioner in the correct diagnosis of a great number of disorders. Blood is found in the gastric contents and in vomitus in conditions associated with erosion of the mucou membranes, in ulcers and in carcinomas. regular and frequent occurrence of occult blood is suggestive of gastro intestinal cancer, gastric or duodenal ulcers or hemorrhoids. In these conditions, the hemorrhage is often so slight that it is not possible to detect blood by microscopic identifications of the erythrocytes (red blood cells) and a sensitive and specific chemical test for occult blood becomes invaluable. In the urine, blood cells (hematuria) or blood pigment hemoglobinuria) is found in typhus, scurvy, purpura, pyemia, nephritis, renal calculi, as the re-,- sult of a burn covering a large part of the body, by the action of various hemolytic toxins, etc.

However, the methods heretofore employed for the detection of occult blood require some technical ability, experience and fresh supplies of reagents which are not always available to the average physician. Thus, the benzidine reaction which is capable of great accuracy and is one of the most delicate tests for the detection of occult blood, requires that a fresh solution of benzidine in glacial acetic acid be prepared daily. Since these solutions change rapidly, especially in contact with light, they rapidly lose their sensitivity and must be discarded. (Journal of Biological Chemistry, volume 19, page 445, 1914; Zentralblatt fur Chirurgie, volume 41, No. 28, 1914.)

Ruttan and Hardisty (Canadian Medical Association Journal, Nov. 1912; Biochemical Bulletin, volume 2, page 225, 1913) have described a modification of the benzidine reaction, using o-tolidine dissolved in glacial acetic acid. This solution is said to remain stable for one month. It has, however, been my experience that after one week, the solution of o-tolidine becomes too In the feces, the

Application September 26, 1940, Serial No. 358,480

insensitive for use since it will no longer detect traces of occult blood that will give a definite positive reaction with a fresh o-tolidine solution.

The benzidine reaction for the detection of occult blood is based, in general, on the reaction of the peroxidase present in the blood with the hydrogen peroxide or the reagent to form active" oxygen. The benzidine acts as an oxygen acceptor and is readily oxidized to a colored quinonoid form. In general, any compound of the 7 general formula:

mN o r Ni where A, B, C, D, E, F, G and H are members of the group consisting of hydrogen and alkyl radicals, may be used as an oxygen acceptor in the benzidine reaction. Examples of such members are benzidine, orthotolidine, metatoli dine, etc.

The present invention has for its further ob ject to provide a means whereby an unskilled tion which remains stable and sensitive over long period of time under all ordinary conditions of temperature and humidity. The basis of the present invention is the finding that such compositions of high sensitivity, good specificity and great stability can be obtained by mixing, in the dry state:

(a). A compound of the general formula:

HzN NP:

where A, B, C, D, E, F, G and H are members of the group consisting of hydrogen and alkyl radicals.

(b) A member a of the group comprising the peroxides, perborates and persulfates ofthe alwithout the use of i .sodium bisulfate, etc., etc. Members 01' this group, when reacted in aqueous solution with a member of the group consisting of the peroxides,

perborates and persulfates of the alkali metals and the alkali-earth metals, will form hydrogen peroxide at a slow and steady rate.

To determine qualitatively whether occult blood is present in a specimen, one drop of the material to be tested (in the case of feces-an aqueous suspension of the specimen) is deposited on a measured quantity of thedry composition, e. g. a five-grain tablet. In the presence of blood, a blue or green colored spot will form within a short time. If, after five minutes, there is no change of color, the specimen is free 01' occult blood. Alternatively, the dry composition may be suspended in water and mixed with the material to be tested. In the presence of occult blood, a blue or green color will be obtained within five minutes.

A semi-quantitative estimation of the amount of occult blood present in the specimen may be made by measuring the time required for the first definite blue or green coloration to be observable. A definite coloration within thirty seconds is reported as four plus, thirty seconds to one minutethree plus, one minute to two minutestwo plus, two minutes to three minutes-one plus,

three minutes to five minutes-plus minus. If

no definite coloration is obtained after five minutes, the test is reported as negative.

' It is obvious, of course, that diluents, excipients, dispersing agents, auxiliary substances and coating materials may be incorporated into these compositions without changing the basis of the present invention. Diluents such as talc, kaolin and gypsum may be added to obtain a tablet with better keeping qualities, or a more stable product. Excipients such as lactose, starch or flour may be added to facilitate the formation of granules. Dispersing agents such as saponin, gelatin, agar, soap, alkali-metal silicates, tragacanth or gum arabic may be added to accelerate the disintegration of the tablet when placed into fluid. Auxiliary agents, such as coloring matters or perfumes may likewise be added. To improve the keeping qualitiesof these compositions in tablet form, they may be covered with a water-dispersible varnish or wax. These compositions may be used in the form or powder, or granules, or dispensed in capsules, or they may be pressed into tablets of such size is convenient for the peri'ormance 01! a sin le te The following examples of typical compositions covered by this invention are intended to define and illustrate but in no way to limit this invention to the reagents, proportions or conditions described therein. Obvious modifications will occur to any *person skilled in the art.

Example I Grams Anhydrous granular citric acid 1000 Benzidine, chemically pure 100 Barium peroxide, heavy powder 250 Talc powder 250 are intimately mixed until homogeneous. The mixture is then compressed into five-grain tablets.

The sensitivity of this composition is such that it will detect one part of blood in one million parts of urine or one and a half million parts of water. Example II Grams Sodium acid pyrophosphate 1000 Orthotolidine, chemically pure 50 Sodium perborate monohydrate 200 Kaolin powder 500 are intimately mixed until homogeneous. The

mixture is then compressed into five-grain tablets.

The sensitivity of this composition is such that it will detect one part of blood in eight hundred thousand parts of urine or one million parts of water.

Having described my invention, what I claim and desire to protect by Letters Patent is:

1. Diagnostic compositions comprising in dry solid form a member of the group consisting oi benzidine, ortho-tolidine and metatolidine, a member of the group consisting of the peroxides, perborates and persulfates of the alka i metals and the alkali-earth metals and a solid compound which contains at least one available acidic hydrogen atom.

2. Diagnostic compositions comprising in dry solid form a member of the group consisting of benzidine, ortho-tolidine and meta-tolidine, barium peroxide and a solid compound which contains at least one available acidic hydrogen atom.

3. Diagnostic compositions comprising in dry solid form a member of the group consisting of benzidine, ortho-tolidine and meta-tolidine, barium peroxide and citric acid.

4. Diagnostic compositions comprising in dry solid form benzidine, barium peroxide and citric acid.

5. Diagonstic compositions comprising in dry solid form orth-tolidine, sodium perborate and sodium acid pyrophosphate.

6. A process for testing for blood in fluids which consists in adding a measured amount or said fiuid to a measured amount of a composition comprising in dry solid form a member of the group consisting of benzidine, ortho-tolidine and meta-tolidine, a member of the group consisting of the peroxides, perborates and persulfates of the alkali metals and the alkali-earth metals, and a solid compound which contains at least one available acidic hydrogen atom, whereby a detectable color-change results.

7. A process for testing for blood in fluids which consists in mixing a measured amount of said fluid with a suspension in water of a measured amount of a composition comprising in dry solid form a member of the group consisting of benzidine, ortho-tolidine and meta-tolidine, a member of the group consisting of the peroxides, perborates and persuliates of the alkali metals and the alkali-earth metals, and a solid compound which contains at least one available acidic hydrogen atom, whereby a detectable color-change results.

JONAS KAMLET.

Referenced by
Citing PatentFiling datePublication dateApplicantTitle
US2436814 *Jun 8, 1943Mar 2, 1948James L LeitchOrtho-tolidine tablet composition
US2569663 *Dec 31, 1947Oct 2, 1951Dearborn Chemicals CoComposition for and method of determining chromate concentrations
US2606101 *Dec 3, 1946Aug 5, 1952Allied Chem & Dye CorpProcess and apparatus for promptly detecting chlorine gas leaks from chlorine containers
US2606102 *Dec 3, 1946Aug 5, 1952Allied Chem & Dye CorpTest strips for detecting low concentrations of chlorine in air
US2676874 *Aug 30, 1950Apr 27, 1954Devine John BerchmansMethod of and means for the rapid approximate quantitative estimation of chloride concentration in aqueous liquids
US2799660 *Apr 15, 1954Jul 16, 1957Miles LabBlood test composition and method
US2838377 *Sep 10, 1953Jun 10, 1958Miles LabBlood test
US2905594 *Apr 25, 1956Sep 22, 1959Herman J MorrisMeans for detecting enzyme activity
US2907638 *Sep 24, 1956Oct 6, 1959Robert L DryerApparatus for quantitative isothermal distillation
US3252762 *May 4, 1961May 24, 1966Miles LabStabilized occult blood diagnostic
US4132527 *Jul 26, 1976Jan 2, 1979Shionogi & Co., Ltd.Diagnostic compositions, diagnosing instruments, and methods of manufacturing same
US4148611 *Jun 28, 1978Apr 10, 1979Miles Laboratories, Inc.Test composition, device and method for the detection of peroxidatively active substances
US4175923 *Jun 26, 1978Nov 27, 1979Friend William GMethod and apparatus for occult blood testing in the home
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US4676950 *Feb 3, 1984Jun 30, 1987Foster Research CorporationIndicator and test device for detecting occult blood
US4939097 *Dec 24, 1987Jul 3, 1990Litmus Concepts, Inc.Fecal occult blood test methods
US4956300 *Oct 16, 1984Sep 11, 1990Helena Laboratories CorporationAid for determining the presence of occult blood, method of making the aid, and method of using the aid
US5053342 *May 7, 1990Oct 1, 1991Litmus Concepts, Inc.Fecal occult blood test reagents
US5081040 *Jun 6, 1989Jan 14, 1992Helena Laboratories CorporationComposition and kit for testing for occult blood in human and animal excretions, fluids, or tissue matrixes
US5196167 *May 9, 1991Mar 23, 1993Helena Laboratories CorporationFecal occult blood test product with positive and negative controls
US5217874 *May 9, 1991Jun 8, 1993Helena Laboratories CorporationFecal occult blood test product with positive and negative controls
US5273888 *Apr 29, 1988Dec 28, 1993Helena Laboratories CorporationChemical test kit and method for determining the presence of blood in a specimen and for verifying the effectiveness of the chemicals
US5702913 *Jun 12, 1989Dec 30, 1997Helena Laboratories CorporationChromgen-reagent test system
US20040214244 *Apr 25, 2003Oct 28, 2004Idexx Laboratories, Inc.Detection of analytes in fecal samples
Classifications
U.S. Classification436/66, 436/904
International ClassificationG01N33/72
Cooperative ClassificationG01N33/725, Y10S436/904
European ClassificationG01N33/72B4