Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
  • G01N33/728—Bilirubin; including biliverdin
• • Y—GENERAL 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
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S436/00—Chemistry: analytical and immunological testing
  • Y10S436/903—Diazo reactions
• • Y—GENERAL 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
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T436/00—Chemistry: analytical and immunological testing
  • Y10T436/14—Heterocyclic carbon compound [i.e., O, S, N, Se, Te, as only ring hetero atom]
  • Y10T436/145555—Hetero-N
  • Y10T436/146666—Bile pigment

Definitions

• This invention relates to a stable test device which undergoes a rapid color change when contacted by a diazo couplable compound.
• the invention also relates to a method for preparing and using such a test device. More particularly, this invention relates to a stable test device comprising a carrier incorporating the dry residue of a diazonium salt solution and to a process for preparing said solution.
• Ictotest which is in the form of a dry tablet, for determination of bilirubin as described in US. Pat. No. 2,854,317.
• the tablet comprises a diazonium salt, sulfosalicylic acid and sodium bicarbonate.
• Ictotest reagent tablet is then placed in the center of the moistened area and two drops of water flowed over the tablet. If bilirubin is present, a color reaction occurs on the mat within about 30 seconds.
• Test devices of the dip-and-read type which combine specialized rapid reacting reagents with a suitable carrier, are widely utilized, particularly in the medical field. Such devices are commonly used for determining pH and de- "ice tecting glucose, protein and the like in body fluids.
• the known test systems for detecting bilirubin were combined with a suitable carrier to form a dip-andread type device, the resulting device was unstable and yielded unreliable results.
• Another object of this invention is to provide a method for preparing a unitary test device that may be used in the detection of bilirubin in a solution.
• This invention is embodied in a novel stable test device which undergoes a color change when contacted by a diazo couplable compound.
• This device comprises a carrier impregnated with a dry residue of a diazonium salt solution comprising a reaction product formed by combining an organic polar solvent, a couplable arylamine, water, a soluble nitrite, a stabilizer and a sulfonic acid.
• This invention is also embodied in a process for preparing the test device, comprising contacting a preferably absorbent carrier with a diazonium salt solution and drying the carrier.
• a polar solvent is used.
• the composition is made up of water and an organic polar solvent such as an alcohol, a ketone, an ester and the like.
• an alcohol such as methanol, ethanol and 2-propanol is used in a ratio of at least about four parts organic solvent to one part water.
• the ratio of organic solvent to water is selected so that there is substantially no precipitate in the diazonium salt solution formed.
• a high ratio of a rapidly evaporating organic solvent is desirable to reduce drying time of a carrier contacted with the solution.
• a couplable arylamine that may be diazotized is beneficially added to the solvent first.
• Such an arylamine is advantageously selected from 2,4-dichloroaniline; p-nitroaniline; p-chloroaniline; 2,5-dichloroaniline; 4-chloro-oanisidine; 3,3'-dimethoxybenzidine; and 2-methoxy-5-nitroaniline.
• Other such arylamines that are capable of forming diazonium compounds may also be used and are considered within the scope of this invention.
• a suitable soluble nitrite that will form nitrous acid in an aqueous acid medium is then added to the solution.
• the soluble nitrite is sodium nitrite, potassium nitrite or calcium nitrite.
• the solution also includes a stabilizer compound.
• a stabilizer compound Such stabilizer has been found beneficial in that any precipitate that appears upon the formation of the diazonium salt forms as a fine precipitate which is readily dissolved. Additionally, the stabilizer has been found to add chemical stability to the final test device.
• the stabilizer may be selected from a broad group of compounds, such as, aromatics or aliphatic sulfonic acid salts, sulfates or sulfonates. Examples of such stabilizers include 1,5-naphthalenedisulfonic acid, disodium salt; Z-naphthalenesulfonic acid, sodium salt; 4,4'-diamino 2,2 biphenyldisulfonic acid, disodium salt and sodium lauryl sulfate.
• An organic acid preferably an organic sulfonic acid, is included in the composition to provide the proper pH for the formation of a diazonium compound and the subsequent coupling reaction of this diazonium compound with the couplable compound.
• the organic sulfonic acid utilized beneficially may be selected from sulfosalicylic acid, sulfamic acid, hexamic and p-toluenesulfonic acid and the like. It is understood that other known sulfonic acids and organic acids may be used within the scope of this invention.
• reagents a polar solvent, an arylamine, water, soluble nitrite, a stabilizer and a sulfonic acid are combined to form the desired diazonium salt solution.
• these reagents are combined in the order in which they have been mentioned, although the order of combination of the polar solvent, arylamine, water, soluble nitrite and stabilizer is not considered critical.
• the second stabilizer may be added after the sulfonic acid.
• Diazo dyes have been prepared using a reagent system similar to that described above.
• the diazo dyes prepared in the prior art systems formed as large particle precipitates which were desirable for their non-dusting properties. Such compounds Were formed at reduced temperatures, about C.
• a diazonium salt solution may be formed at room temperature (about 23 C.) and retained in a solution which is substantially free of precipitate at such temperature with reagents similar to those of prior art systems.
• a carrier is contacted with this diazonium salt solution and such carrier is then dried, the residue of the diazonium salt solution forms in small particles that are readily retained by the carrier.
• the proportions of the reagents used to form the diazonium salt solution are preferably within the following ranges which are expressed in weight/ volume percent, as appropriate:
• the ratio of solvent to water is maintained at 4:1 or higher to retain the diazonium salt in solution and promote the drying of the carrier.
• the weight ratio of acid to arylamine compound is preferably maintained at about :1.
• a suitable carrier may be utilized that is capable of retaining the dry residue of the diazonium salt solution incorporated with the carrier.
• This carrier is preferably mounted in such a manner that it may be readily contacted by a fiuid suspected of including the couplable compound to be detected.
• the carrier for the test device may be an integral part of a larger portion serving as a handle or may be a separate part that is affixed to a larger handle or handling device.
• Such a carrier may l comprise an absorbent material that is dipped in or otherwise contacted with the diazonium salt solution and subsequently dried.
• Suitable carriers include, for example, a porous material such as filter paper, glass fiber paper,
• the carrier may also comprise I a polymeric gel which has been premixed with the test solution and dried so as to form a semipermeable membrane structure.
• the gel may be formed by solvent evaporation and may include a cross-linking agent.
• test device is not considered critical so long as a suitable means is provided for contacting the area including the dry residue of the diazonium salt solution with the fluid to be tested.
• a test device may also be prepared by impregnating a piece of filter paper, which functions as a carrier, with the diazonium salt solution and drying the same.
• the impregnated paper is preferably dried at a moderate temperature of about 65 to 70 C. in moving air.
• the temperature is not elevated excessively as this is believed to have a deleterious effect upon the diazonium compounds.
• the dried carrier may then be divided into small portions and affixed to a larger handle for easier handling.
• EXAMPLE 1 A solution was prepared by combining in a glass vessel, in the order listed and with continuous mixing, the following compounds:
• the compounds when combined were at ambient room temperature (about 23 C.) and the temperature of the solution remained at about ambient room temperature without additional temperature control throughout the formation of the solution.
• EXAMPLE 2 A solution was prepared as described in Example 1 except that 0.05 g. of 2,4-dichloroaniline and 0.5 g. of Sulfosalicylic acid were used. This solution was observed to have substantially the same properties as the solution of Example 1.
• EXAMPLE 3 A solution was prepared as described in Example 1 except that 0.5 g. of 2,4-dichloroaniline and 5.0 g. of sulfosalicylic acid were used. This solution was observed to have substantially the same properties as the solution of Example 1.
• EXAMPLE 11 A solution was prepared as described in Example 1 except that hexamic acid was used in place of Sulfosalicylic acid. The resulting solution was observed to have substantially the same properties as the solution of Example 1.
• EXAMPLE 12 A solution was prepared as described in Example 1 except that sulfamic acid was used in place of sulfosalicylic acid. The resulting solution was observed to have substantially the same properties as the solution of Example 1.
• EXAMPLE 13 A solution was prepared as described in Example 1 except that p-toluenesulfonic acid was used in place of sulfosalicylic acid. The resulting solution was observed to have substantially the same properties as the solution of Example 1.
• EXAMPLE 14 A strip of filter paper (Eaton and Dikeman No. 641) was immersed in the solution of Example 1 and immediately removed. This paper was dried between about and C. for between about 8 and 10 minutes with air movement over the surface of the paper. The dried paper had a cream white color.
• Bilirubin test solutions were prepared from a pathological urine sample obtained from a patient showing signs of jaundice. The urine obtained from the patient was treated with Ictotest and a very large reaction was observed which indicated a high concentration of bilirubin. This urine was also assayed by a modification of the E. G. Godfried procedure for bilirubin described in Biochem. J 28, 20562060, 1934 and found to have approximately 1 mg. percent bilirubin. The pathological urine was further diluted With normal urine to concentrations of 0.5, 0.25 and 0.1 mg. percent. Each solution was tested with test devices of this invention as prepared above and gradations of pink were observed. A deeper pink, approaching a pink-lavender, was observed for the solutions of higher concentrations.
• EXAMPLE 15 Strips of glass fiber paper (Gelman Type A-1306 and Type E-748) were processed in place of filter paper according to the procedure of Example 14. The resulting dried glass fiber paper had an appearance substantially the same as the paper of Example 14. When contacted by the test solutions of Example 14 color changes as noted in Example 14 were observed.
• Example 16 In this example the procedure of Example 14 was followed except that a polypropylene felt (Polypropylene MDSE. No. Po-8110 by American Felt Company), was used in place of filter paper. The dried matrix had substantially the same color appearance and color reaction to the test solutions as observed in Example 14.
• a polypropylene felt Polypropylene MDSE. No. Po-8110 by American Felt Company
• EXAMPLE 17 A solution was prepared according to the proportions of Example 1.
• a 4-inch wide sheet of Eaton and Dikeman No. 641 filter paper was passed through this solution and dried between about 65 and 70 C. for about 10 minutes with circulating air. After drying, the paper was contacted with one pressure sensitive adhesive surface of a double-faced pressure sensitive cellulosic fiber reinforced adhesive tape, slit longitudinally into strips each inch wide and wound into rolls. A roll was then mounted on an axis and a wax paper backing or liner was removed from the outer adhesive surface of the double-faced pressure sensitive tape bonded to the paper. The exposed surface of the pressure sensitive tape was continuously bonded for the length of the roll to a surface of a 3% inch wide sheet of polystyrene film 0.01 inch thick. The pressure sensitive tape was securely bonded to the film. The laminate was cut across the length of the strip into strips /5 inch long. The device so formed was contacted with the test solutions of Example 14 and a color change was noted as in Example 14.
• a stable test device which exhibits a color change when contacted by a diazo couplable compound, said device comprising a carrier incorporating a dry residue of a stable diazonium salt solution comprising a reaction product formed by combining an organic polar solvent, an arylamine capable of forming a diazonium salt, water, a soluble nitrite, a sulfonic acid and a stabilizer which is selected from aromatic or aliphatic sulfonic acid salts, sulfates or sulfonates.
• a stable test device according to claim 1 in which the carrier is an absorbent material.
• a stable test device in which the polar solvent is an alcohol, an ether, a ketone or an ester.
• a stable test device in which the arylamine is 2,4-dichloroaniline, p-nitroaniline, pchloroaniline, 2,5-dichloroaniline, 4-chloro-o-anisidine, 3,3-dimethyoxybenzidine, or 2-methoxy-5-nitroaniline.
• a stable test device in which the soluble nitrite is sodium nitrite, potassium nitrite or calcium nitrite.
• aromatic sulfonic acid salt is a member selected from the group consisting of 1,5-napthalenedisulfonic acid, disodium salt; 2-napthalenesulfonic acid, sodium salt; and 4,4-diamino-2,2'-biphenyldisulfonic acid, disodium salt.
• a stable test device in which the stabilizer is sodium lauryl sulfate.
• sulfonic acid is a member selected from the group consisting of sulfosalicylic acid, sulfamic acid, and hexamic acid.
• a stable test device in which the polar solvent is methanol; the arylamine is 2,4-dichloroaniline; the soluble nitrite is sodium nitrite, the stabilizers are 1,5-napthalenedisulfonic acid, disodium salt and sodium lauryl sulfate and the sulfonic acid is sulfosalicylic acid.
• a stable test device in which the carrier is an absorbent material; the polar solvent is an alcohol, ether, ketone or ester; the arylamine is 2,4-dichloroaniline, 4,4'-methylenebis (N,N-dimethylaniline), pnitroaniline, p-chloroaniline, 2,5-dichloroaniline, 4-chloroo-anisidine, 3,3'-dimethoxybenzidine, or 2-methoxy-5- nitroaniline; the soluble nitrite is sodium nitrite; the stabilizer is 1,5-napthalenedisulfonic acid, disodium salt; Z-napthalene sulfonic acid, sodium salt, 4,4'diamino-2,2'- biphenyldisulfonic acid, disodium salt or sodium lauryl sulfate and the sulfonic acid is sulfosalicylic acid, sulfamic acid or hexamic acid.
• a process for preparing the test device of claim 1 comprising mixing in the order stated the polar solvent, the arylamine, the soluble nitrite, the stabilizer and the sulfonic acid to form the stable diazonium salt solution, contacting a carrier with the diazonium salt solution and drying the carrier leaving a dry residue of the diazonium salt solution thereon.
• a stable test device in which the arylamine is selected from aniline, substituted aniline, diaminobiphenyl or substituted diaminobiphenyl.
• a process for detecting a diazo couplable compound in a solution comprising contacting the test device of claim 1 with the solution and observing the color change of the test device.
• a process according to claim 13 in which the diazo couplable compound to be detected is bilirubin in an aqueous solution.

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Cited By (14)

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• 1969
  • 1969-02-17 US US799928A patent/US3585001A/en not_active Expired - Lifetime
  • 1969-12-23 ES ES374872A patent/ES374872A1/es not_active Expired
• 1970
  • 1970-02-03 GB GB5174/70D patent/GB1298762A/en not_active Expired
  • 1970-02-04 CH CH161070A patent/CH533837A/de not_active IP Right Cessation
  • 1970-02-05 NL NL7001638A patent/NL7001638A/xx unknown
  • 1970-02-13 BR BR216743/70A patent/BR7016743D0/pt unknown
  • 1970-02-13 NO NO00504/70A patent/NO128970B/no unknown
  • 1970-02-16 DK DK74170AA patent/DK134872B/da unknown
  • 1970-02-16 DE DE2007013A patent/DE2007013B2/de active Pending
  • 1970-02-16 JP JP45012668A patent/JPS4929475B1/ja active Pending
  • 1970-02-16 SE SE7001954A patent/SE374821B/xx unknown
  • 1970-02-16 FR FR7005475A patent/FR2035492A5/fr not_active Expired
  • 1970-02-17 BE BE746099D patent/BE746099A/xx unknown

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