US3418083A - Hydrophobic test strip for analyzing whole blood - Google Patents

Hydrophobic test strip for analyzing whole blood Download PDF

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Publication number
US3418083A
US3418083A US53501066A US3418083A US 3418083 A US3418083 A US 3418083A US 53501066 A US53501066 A US 53501066A US 3418083 A US3418083 A US 3418083A
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Prior art keywords
whole blood
agent
impregnated
blood
test
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Expired - Lifetime
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Rey Hans-Georg
Rieckmann Peter
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Roche Diagnostics GmbH
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Roche Diagnostics GmbH
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/54Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving glucose or galactose
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • 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
    • G01N33/52Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G01N33/521Single-layer analytical elements
    • 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
    • Y10STECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00Chemistry: molecular biology and microbiology
    • Y10S435/805Test papers
    • 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
    • Y10T436/14Heterocyclic carbon compound [i.e., O, S, N, Se, Te, as only ring hetero atom]
    • Y10T436/142222Hetero-O [e.g., ascorbic acid, etc.]
    • Y10T436/143333Saccharide [e.g., DNA, etc.]
    • 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
    • Y10T436/14Heterocyclic carbon compound [i.e., O, S, N, Se, Te, as only ring hetero atom]
    • Y10T436/142222Hetero-O [e.g., ascorbic acid, etc.]
    • Y10T436/143333Saccharide [e.g., DNA, etc.]
    • Y10T436/144444Glucose

Description

United States Patent 3,418,083 HYDROPHOBIC TEST STRIP FOR ANALYZING WHOLE BLOOD Hans-Georg Rey, Mannheim, and Peter Rieckmann, Mannheim-Waldhof, Germany, assignors to C. F. Boehringer & Soehne G.m.b.H., Mannheim-Waldhof, Germany No Drawing. Filed Mar. 17, 1966, Ser. No. 535,010 Claims priority, application Germany, May 20, 1965, B 82,023 8 Claims. (Cl. 23-253) This invention relates to a diagnostic agent for the detection of biochemical and chemical components in whole blood and more particularly relates to a test strip for use in the detection of chemical and biochemical components in whole blood, a process for the preparation of such test strips and a method for using the same.

The detection of biochemical and chemical components in whole blood has hirtherto been a very laborious, expensive and time-consuming procedure which can only be carried out in suitably equipped laboratories and with skilled personnel. The whole blood must first be processed to the serum or plasma in which only then can be analyzed for the substances of interest. Whole blood is not suitable as the strong coloring properties of hemoglobin would interfere or render inaccurate all color reactions.

Recently, procedures have been described which make possible the detection of glucose or urea in whole blood (of. US. Patent No. 3,092,465 and British Patent No. 922,665). The principle of these new procedures involves the covering of a test paper strip on the outside thereof with a semipermeable wall (made from various cellulose derivatives). When whole blood is placed on a strip of this type, the blood fluid, together with the components dissolved therein, passes through the semi-permeable film giving rise to a color reaction in the test paper, the red blood corpuscles remaining behind on the film. After a predetermined period of time, the blood coagulum (red cells) must be washed off in order that the color reaction developed in the underlying paper can be seen.

It is an object of the present invention to provide a new and improved test paper strip for use in detecting chemical and biochemical components of whole blood.

A further object of the invention is to provide a new and improved test paper strip for use in detecting chemical and biochemical components of whole blood characterized by the extreme ease with which they are produced and used.

These and other objects and advantages will be apparent from a consideration of the following disclosure.

In accordance with the invention, it has now been found that diagnostic agents for the detection of biochemical and chemical components in whole blood which are outstandingly useful and can be prepared in a substantially simpler and more economic manner than possible heretofore, are obtained by impregnating an absorbent carrier (preferably filter paper) impregnated with specific reagents and indicators which absorbent carrier is prior to, during, or following the impregnation hydrophobed. This result is completely surprising. It was not to have been foreseen that a hydrophobed absorbent carrier such as hydrophobed filter paper would be moistened with serum or plasma to such an extent that the reaction can take place in the filter paper While the red blood corpuscles are prevented, by the hydrophobing, from penetrating into 3,418,083 Patented Dec. 24, 1968 the paper so that they can be completely rinsed ofi' or wiped off with gauze, cloth, wadding or by cotton wool. In the case of non-hydrophobed test paper strips, the red blood corpuscles penerate so deeply into the filter paper that they can be neither wiped off nor rinsed out. However, due to their intense inherent color, they interfere with the actual color reactions to such an extent that the color change can no longer be observed or appreciated.

Exemplary instances of agents which can be used for the hydrophobing of the absorbent carriers are natural and synthetic fats, oils and waxes, as well as, and preferably, paraflins, silicones and mixtures thereof. Generally,

the conventional test paper strips can be hydrophobed following impregnation with the indicator solutions. However, dependable and satisfactory test paper strips are also obtained when the indicator dyestuff, together with the hydrophobing agent, is employed as impregnant being applied onto test paper strips which have already been impregnated with the other reagents.

For the production of the hydrophobed diagnostic agents according to the present invention, there can be used not only filter paper, but also other absorbent carriers, such as asbestos paper and Wood rodlets. However, the pores of the wood can sometimes be disturbing since red blood corpuscles collect therein and can only be wiped off or rinsed out with difficulty.

The diagnostic agents according to the present invention can be utilized for all of the clinical tests which are of importance in the analysis of blood, especially for the clinical screening tests, as for example, the determination of glucose, galactose, phenyl-alanine, serum cholinesterase, urea and uric acid, as well as for phosphatases, transaminases, dehydrogenases, trypsin, histidine, and the like.

Furthermore, an important advantage of the hydrophobed diagnostic agents according to the invention, in comparison with all diagnostic agents which have previously been suggested or used for the investigation of whole blood, is the extreme ease with which they are produced and used since sensitive and expensive special coatings or sealings are not necessary.

The following examples are given for the purpose of illustrating the present invention and are not to be construed as a limitation thereof.

EXAMPLE 1 Production of a test strip suitable for the detection of glucose in whole blood.

impregnation solution I Grams Peroxidase 0.012 Glucose oxidase 1.300 Primary potassium phosphate 0.269 Secondary sodium phosphate 0.004 Glycocoll 6.000 Distilled water ad 100.0 ml.

Impregnation solution II Grams o-Tolidine 0.420 Paraffin 4.000

Methylene chloride ad 100.0 ml.

Schleicher and Schiill filter paper No. 2312 was successively impregnated with impregnation solutions I and II, dried and cut up into strips of mm. width.

When a strip thereby obtained was moistened with a drop of blood, taken, for example, from a finger or ear lobe, the blood left to act upon the strip for 1 minute and the blood then rinsed off with water, there resulted a blue color reaction which was dependent upon the glucose content of the blood and which could be evaluated by reference to a comparative color scale. The same results were obtained when the methylene chloride was replaced by petrol ether.

EXAMPLE 2 Production of a test strip suitable for the detection of galactose in whole blood.

Impregnation solution I Methylene chloride ad 100.0 ml.

Schleicher and Schiill filter paper No. 2312 was successively impregnated with impregnation solutions -I and II, dried and cut up into strips. Blood containing galactose obtained as described above was applied to the test strips in a manner analogous to that described in Example 1 and the strips further processed as therein set forth. The test strips evidence the galactose content of the blood by a blue color reaction, the intensity of which is directly related to the amount of galactose present.

EXAMPLE 3 In place of the impregnation solution II used in Example 1, there was used a solution having the following composition:

Grams o-Tolidine 0.420 Silicone impregnation agent 2.000

Methylene chloride ad 100.0 m1.

Apart from this difference, the test strip was produced in exactly the same way as described in Example 1. The test strips thereby obtained had the same properties as those of Example 1 and produced the same results.

EXAMPLE 4 Phenyl-alanine test paper Filter paper was impregnated with L-amino acid oxidase, peroxidase and o-tolidine, together with a 0.2 molar phosphate buffer having a pH of 6.5. The dried paper was subsequently impregnated with a solution of 4% paraflin in methylene chloride. The phenyl-alanine test paper obtained in this manner demonstrated, upon the application thereto of blood, containing phenylalanine in a manner analogous to that described in Example 1, the content of L-phenyl-alanine by a blue color reaction which could be quantitatively evaluated by reference to a color table.

The same results were obtained when the paraffin was replaced by 2% silicone.

We claim:

1. A diagonstic agent for use in the detection of chemical and biochemical components in whole blood comprising an absorbent carrier support impregnated with a composition forming color in response to said chemical component and treated with a hydrophobing agent rendering the impregnated absorbent carrier hydrophobic, wherein said hydrophobing agent is a member selected from the group consisting of natural and synthetic fats, oils, waxes, parafiins, silicones, and mixtures thereof.

2. A diagnostic agent according to claim 1, wherein said absorbent carrier support is a member selected from the group consisting of filter paper, asbestos paper, and wood rodlets.

3. A diagonstic test agent according to claim 1, wherein said absorbent carrier support is impregnated with the following composition for forming color in response to said chemical component:

Grams Peroxidase 0.012 Glucose oxidase 1.300 Primary potassium phosphate 0.269 Secondary sodium phosphate 0.004 Glycocoll 6.000

Distilled water ad 100.0 ml. and is hydrophobed with the following composition:

Grams o-Tolidine 0.420 Parafiin 4.000

Methylene chloride ad 100.0 ml.

4. A diagnostic test agent according to claim 1, wherein said absorbent carrier support is impregnated with the following composition for forming color in response to said chemical component:

Grams Peroxidase 0.012 Galactose oxidase 0.300 Primary potasium phosphate 0.746 Secondary sodium phosphate 2.585

Distilled water ad 100.0 ml. and is hydrophobed with the following composition:

Grams o-Tolidine 0.420 Parafiin 4.000

Methylene chloride ad 100.0 ml.

5. A diagnostic test agent according to claim 1, wherein said absorbent carrier support is impregnated with the following composition for forming color in response to said chemical component:

Grams Peroxidase 0.012 Glucose oxidase 1.300 Primary potassium phosphate 0.269 Secondary sodium phosphate 0.004 Glycocoll 6.000

Distilled water ad 100.0 ml.

and is hydrophobed with the following composition:

Grams o-Tolidine 0.420 Silicone impregnation agent 2.000

Methylene chloride ad 100.0 ml.

6. The method of providing diagnostic agents for use in the detection of chemical and biochemical components in whole blood comprising impregnating an absorbent carrier support with a composition forming color in response to said chemical component and thereafter treating said impregnated carrier support with a hydrophobic agent rendering the impregnated carrier support hydrophobic, wherein said hydrophobic agent is a member selected from the group consisting of natural and synthetic fats, oils, waxes, paraffins, silicones and mixtures thereof.

7. Method according to claim 6, wherein said composition for forming color in response to said chemical component comprises a mixture of specific reagents and indicators.

8. Method according to claim 6, wherein said composition for forming color in response to said chemical component comprises a mixture of specific reagents and indicators and wherein said absorbent carrier is first 3,418,083 5 6 impregnated with said specific reagents and thereafter MORRIS O. WOLK, Primary Examiner. treated with the indicator and hydrophobing agent.

E. A. KATZ, Assistant Examiner. References Cited UNITED STATES PATENTS 5 US. Cl. X.R.

3,092,465 6/1963 Adams et a1. 3,298,789 1/1967 Mast 252408, 195-4035, 167-845

Claims (1)

1. A DIAGONSTIC AGENT FOR USE IN THE DETECTION OF CHEMICAL AND BIOCHEMICAL COMPONENTS IN WHOLE BLOOD COMPRISING AN ABOSRBENT CARRIER SUPPORT IMPREGNATED WITH A COMPOSTION FORMING COLOR IN RESPONSE TO SAID CHEMICAL COMPONENT AND TREATED WITH A HYDROPHOBING AGENT RENDERING THE IMPREGNATED ABSORBENT AGENT IS A MEMBER SELECTED FROM THE GROUP CONSISTING OF NATURAL AND SYNTEHTIC FATS, OILS, WAXES, PARAFFFINS, SILICONES, AND MIXTURES THEREOF.
US3418083A 1965-05-20 1966-03-17 Hydrophobic test strip for analyzing whole blood Expired - Lifetime US3418083A (en)

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DE1965B0082023 DE1598048B2 (en) 1965-05-20 1965-05-20 Diagnostic means for detecting constituents of whole blood

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US (1) US3418083A (en)
BE (1) BE681364A (en)
DE (1) DE1598048B2 (en)
ES (1) ES326843A1 (en)
FI (1) FI41879C (en)
FR (1) FR1484845A (en)
GB (1) GB1083683A (en)
LU (1) LU51140A1 (en)
NL (2) NL6606814A (en)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3607093A (en) * 1968-02-15 1971-09-21 Schering Corp Devices for testing biological liquids
US3913393A (en) * 1972-02-25 1975-10-21 Omnium Tech Administrat Emulsion layer of coloured indicators for determining spectra of the effects of a fluid on a surface
US3917453A (en) * 1974-08-16 1975-11-04 Polaroid Corp Method and device for determining the concentration of a substance in a fluid
US4017261A (en) * 1974-10-16 1977-04-12 Lachema, Narodni Podnik Biological diagnostic test strip and method of producing same
US4160008A (en) * 1978-01-26 1979-07-03 Miles Laboratories, Inc. Multilayered test device for determining the presence of a liquid sample component, and method of use
US4234313A (en) * 1974-11-11 1980-11-18 Burroughs Wellcome Co. Device and method for quantitative uric acid testing
EP0054773A1 (en) * 1980-12-15 1982-06-30 Miles Laboratories, Inc. Method and device for detecting glucose concentration
US4385114A (en) * 1974-12-21 1983-05-24 Boehringer Mannheim Gmbh Oxidation indicators comprising 3,3',5,5'-tetraalkylbenzidine compounds
EP0133700A2 (en) * 1983-08-10 1985-03-06 E.I. Du Pont De Nemours And Company Improved multilayered test device for detecting analytes in liquid test samples
US4557900A (en) * 1982-09-28 1985-12-10 Cardiovascular Devices, Inc. Optical sensor with beads
US4689309A (en) * 1985-09-30 1987-08-25 Miles Laboratories, Inc. Test device, method of manufacturing same and method of determining a component in a sample
US4965087A (en) * 1982-12-07 1990-10-23 Avl Ag Method of making a sensor element for fluorescence-optical measurements
US5312759A (en) * 1989-12-20 1994-05-17 Iatron Laboratories, Inc. Method for measurement of fructosamines using 1,2-quinones
WO1998002249A1 (en) * 1996-07-17 1998-01-22 Robert William Cunningham New test device for mass screening
US5962215A (en) * 1996-04-05 1999-10-05 Mercury Diagnostics, Inc. Methods for testing the concentration of an analyte in a body fluid
US20030211006A1 (en) * 2002-05-10 2003-11-13 Suyue Qian Multilayer reagent test strips that include at least one fluid flow control layer and methods for using the same
WO2006000999A1 (en) * 2004-06-24 2006-01-05 University College Cork - National University Of Ireland, Cork Support for biological or chemical arrays having increased spot quality
WO2013045443A1 (en) 2011-09-28 2013-04-04 Emilia Bramanti Measurement of lactic acid in biological fluids

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0146029B2 (en) * 1982-08-02 1989-10-05 Eiken Chemical
CA1271398A (en) * 1985-06-28 1990-07-10 Miles Laboratories, Inc. Diagnostic test device
DE3800661A1 (en) * 1988-01-13 1989-07-27 Zuckerindustrie Verein Test strip consisting of an absorbent material or with an absorbent coating of this material and process for its preparation

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3092465A (en) * 1960-03-25 1963-06-04 Miles Lab Diagnostic test device for blood sugar
US3298789A (en) * 1964-12-14 1967-01-17 Miles Lab Test article for the detection of glucose

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3092465A (en) * 1960-03-25 1963-06-04 Miles Lab Diagnostic test device for blood sugar
US3298789A (en) * 1964-12-14 1967-01-17 Miles Lab Test article for the detection of glucose

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3607093A (en) * 1968-02-15 1971-09-21 Schering Corp Devices for testing biological liquids
US3913393A (en) * 1972-02-25 1975-10-21 Omnium Tech Administrat Emulsion layer of coloured indicators for determining spectra of the effects of a fluid on a surface
US3917453A (en) * 1974-08-16 1975-11-04 Polaroid Corp Method and device for determining the concentration of a substance in a fluid
US4017261A (en) * 1974-10-16 1977-04-12 Lachema, Narodni Podnik Biological diagnostic test strip and method of producing same
US4234313A (en) * 1974-11-11 1980-11-18 Burroughs Wellcome Co. Device and method for quantitative uric acid testing
US4385114A (en) * 1974-12-21 1983-05-24 Boehringer Mannheim Gmbh Oxidation indicators comprising 3,3',5,5'-tetraalkylbenzidine compounds
US4160008A (en) * 1978-01-26 1979-07-03 Miles Laboratories, Inc. Multilayered test device for determining the presence of a liquid sample component, and method of use
EP0054773A1 (en) * 1980-12-15 1982-06-30 Miles Laboratories, Inc. Method and device for detecting glucose concentration
US4557900A (en) * 1982-09-28 1985-12-10 Cardiovascular Devices, Inc. Optical sensor with beads
US4965087A (en) * 1982-12-07 1990-10-23 Avl Ag Method of making a sensor element for fluorescence-optical measurements
EP0133700A2 (en) * 1983-08-10 1985-03-06 E.I. Du Pont De Nemours And Company Improved multilayered test device for detecting analytes in liquid test samples
EP0133700A3 (en) * 1983-08-10 1987-10-28 E.I. Du Pont De Nemours And Company Improved multilayered test device for detecting analytes in liquid test samples
US4689309A (en) * 1985-09-30 1987-08-25 Miles Laboratories, Inc. Test device, method of manufacturing same and method of determining a component in a sample
US5312759A (en) * 1989-12-20 1994-05-17 Iatron Laboratories, Inc. Method for measurement of fructosamines using 1,2-quinones
US5962215A (en) * 1996-04-05 1999-10-05 Mercury Diagnostics, Inc. Methods for testing the concentration of an analyte in a body fluid
US6818180B2 (en) 1996-04-05 2004-11-16 Roche Diagnostics Operations, Inc. Devices for testing for the presence and/or concentration of an analyte in a body fluid
WO1998002249A1 (en) * 1996-07-17 1998-01-22 Robert William Cunningham New test device for mass screening
US20030211006A1 (en) * 2002-05-10 2003-11-13 Suyue Qian Multilayer reagent test strips that include at least one fluid flow control layer and methods for using the same
WO2006000999A1 (en) * 2004-06-24 2006-01-05 University College Cork - National University Of Ireland, Cork Support for biological or chemical arrays having increased spot quality
WO2013045443A1 (en) 2011-09-28 2013-04-04 Emilia Bramanti Measurement of lactic acid in biological fluids
US9234887B2 (en) 2011-09-28 2016-01-12 Power Fit S.R.L. Measurement of lactic acid in biological fluids
US9702848B2 (en) 2011-09-28 2017-07-11 Power Fit S.R.L. Measurement of lactic acid in biological fluids

Also Published As

Publication number Publication date Type
LU51140A (en) application
NL129843C (en) grant
NL6606814A (en) 1966-11-21 application
ES326843A1 (en) 1967-03-16 application
FI41879C (en) 1970-03-10 grant
LU51140A1 (en) 1966-07-18 application
DE1598048B2 (en) 1977-06-16 application
GB1083683A (en) 1967-09-20 application
DE1598048A1 (en) 1969-06-12 application
FI41879B (en) 1969-12-01 application
FR1484845A (en) 1967-06-16 grant
BE681364A (en) 1966-11-21 grant

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