US3653836A - Reagent for determining blood sugar - Google Patents
Reagent for determining blood sugar Download PDFInfo
- Publication number
- US3653836A US3653836A US66953A US3653836DA US3653836A US 3653836 A US3653836 A US 3653836A US 66953 A US66953 A US 66953A US 3653836D A US3653836D A US 3653836DA US 3653836 A US3653836 A US 3653836A
- Authority
- US
- United States
- Prior art keywords
- percent
- weight
- reagent
- blood sugar
- composition
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 51
- 239000008280 blood Substances 0.000 title claims abstract description 36
- 210000004369 blood Anatomy 0.000 title claims abstract description 36
- RNVCVTLRINQCPJ-UHFFFAOYSA-N o-toluidine Chemical compound CC1=CC=CC=C1N RNVCVTLRINQCPJ-UHFFFAOYSA-N 0.000 claims abstract description 36
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims abstract description 30
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 26
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 claims abstract description 22
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 claims abstract description 14
- 150000005215 alkyl ethers Chemical class 0.000 claims abstract description 10
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Natural products NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000000203 mixture Substances 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- KTCBLMXNWYGCRT-UHFFFAOYSA-N 1-[2-(1h-indol-3-yl)ethyl]-3-methylthiourea Chemical compound C1=CC=C2C(CCNC(=S)NC)=CNC2=C1 KTCBLMXNWYGCRT-UHFFFAOYSA-N 0.000 claims description 2
- -1 alkylene glycol Chemical compound 0.000 claims description 2
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 2
- 239000002904 solvent Substances 0.000 abstract description 5
- 208000013016 Hypoglycemia Diseases 0.000 abstract description 3
- 230000002035 prolonged effect Effects 0.000 abstract 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 27
- 229960000583 acetic acid Drugs 0.000 description 13
- 239000012362 glacial acetic acid Substances 0.000 description 12
- 230000035945 sensitivity Effects 0.000 description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 10
- 239000008103 glucose Substances 0.000 description 10
- 238000005259 measurement Methods 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 5
- 230000008033 biological extinction Effects 0.000 description 5
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical group OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000005548 Hexokinase Human genes 0.000 description 2
- 108700040460 Hexokinases Proteins 0.000 description 2
- KKEYFWRCBNTPAC-UHFFFAOYSA-N Terephthalic acid Chemical compound OC(=O)C1=CC=C(C(O)=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000003544 deproteinization Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001771 impaired effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- QQVIHTHCMHWDBS-UHFFFAOYSA-N isophthalic acid Chemical compound OC(=O)C1=CC=CC(C(O)=O)=C1 QQVIHTHCMHWDBS-UHFFFAOYSA-N 0.000 description 2
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Chemical group OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 2
- 239000012429 reaction media Substances 0.000 description 2
- 229960004889 salicylic acid Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- XNWFRZJHXBZDAG-UHFFFAOYSA-N 2-METHOXYETHANOL Chemical compound COCCO XNWFRZJHXBZDAG-UHFFFAOYSA-N 0.000 description 1
- ZNQVEEAIQZEUHB-UHFFFAOYSA-N 2-ethoxyethanol Chemical compound CCOCCO ZNQVEEAIQZEUHB-UHFFFAOYSA-N 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 239000012445 acidic reagent Substances 0.000 description 1
- 150000001323 aldoses Chemical class 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 150000001991 dicarboxylic acids Chemical class 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- DQYBDCGIPTYXML-UHFFFAOYSA-N ethoxyethane;hydrate Chemical compound O.CCOCC DQYBDCGIPTYXML-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 150000002895 organic esters Chemical group 0.000 description 1
- 238000010517 secondary reaction Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000011877 solvent mixture Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
Images
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
- G01N33/66—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood sugars, e.g. galactose
-
- 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]
- Y10T436/142222—Hetero-O [e.g., ascorbic acid, etc.]
- Y10T436/143333—Saccharide [e.g., DNA, etc.]
- Y10T436/144444—Glucose
Definitions
- the present invention is concerned with a reagent for the determination of blood sugar by the o-toluidine method, which reagent does not contain glacial acetic acid.
- isophthalic acid and terephthalic acid are useless for weight, based on total weight of composition.
- the two above-mentioned glacial acetic acid-free reagents overcome the problem of malodor and also somewhat improve the sensitivity of the test.
- This sensitivity is, in the case of the usual normal values for the blood sugar content in whole blood of 60 100 mg. percent, below the value ofE 0.100 at the measurement wavelength of 578 nm usually employed.
- a further disadvantage of the two above-mentioned known reagents is the instability of the solvent mixture which is due to the content of readily volatile components.
- the increased evaporation rate is of significance because the reagent must be heated for 8 minutes in a boiling waterbath for the development of the coloration.
- the dissolved substances start to crystallize out.
- phthalic acid is an outstanding catalyst for An especially preferred composition consists of 0.075 0.15 percent by weight thiourea, 20 25 percent by weight phthalic acid, 35 55 percent by weight dimethyl formamide, 20 40percent by weight ethylene glycol mono-lower alkyl ether and 7.5 10 percent by weight o-toluidine, based on total weight of composition.
- lower alkly is to be understood to mean an alkyl radical containing up to six carbon atoms.
- the reagent according to the present invention can contain up to about 5 percent water, without its usefulness being thereby impaired.
- the reagent according to the present invention does not result in any unpleasant smell nor is there any danger of a loss of solvent when heating in order to bring about formation of the colored material.
- the reagent is colorless to pale yellow in color and the viscosity is low so that there is no impairment of the pipetting due to air bubbles or after-runnings.
- the reagent is also storage stable although, in the case of comparatively long periods of storage at an elevated temperature (33 C. an intensification of the yellow color can occur. However, this influences neither the measurement sensitivity to any considerable extent nor the stability of the color formed.
- FIG. 1 is a plot extinction values against glucose concentration
- the superior sensitivity of the reagent according to the present invention is of great practical importance since it is hereby possible, even in the cases of hypoglycaemia, as well as in the cases in which only very little blood is available, i.e., in the case of blood sugar measurement values of 30 50 mg. percent, to obtain extinctions of the order of 0.050 to 0.082 at 578 nm, measured against a blank, so that errors of measurement in the reading off of the photometer are very considerably reduced.
- a further advantage of the reagent according to the present invention lies in the fact that in the region of the especially preferred composition, it represents a so-called optimum composition. This means that all individual components of the mixture are present in the concentration in which the sensitivity, the period of incubation of the test and the stability of the resultant color have their maxima and a slight alteration of the conditions does not cause any significant differences in the course of the test or in the measurement operation. The working errors are, therefore, in the case of this composition, extremely low.
- the color which arises in the case of the o-toluidine reaction is, in principle, unstable. It is assumed that an unknown secondary reaction follows by means of which the extinction at 640 nm, the maximum, and at the measurement wavelength of 578 nm, decreases in the course of time. This instability of the color is reduced in the case of the reagent according to the had at ambient temperature at the end of the incubation.
- glycollic acid reagent was used as comparison reagent.
- superior stability of the reagent according to the present invention This improvement represents an important advantage when a large series of tests are to be measured one after the other.
- a further advantage of the reagent according to the present invention lies in the fact that the stability is independent of the glucose concentration in the sample. The possibilities of error of the method are hereby minimized.
- a further advantage of the reagent according to the present invention is its cheapness.
- the cost of phthalic acid is only about one-ninth of the cost of glycollic acid.
- Example 1 ml. ethylene glycol monoethyl ether were then added thereto and finally 18.0 ml. o-toluidine was stirred in.
- 0.1 ml. of blood was pipetted into 1.0 ml. 3 percent trichloroacetic acid, mixed and, after standing for a few minutes, centrifuged.
- Areagent composition for determination of blood sugar which composition comprises o-toluidine, thiourea, an ethylene glycol mono-lower alkyl ether, phthalic acid, and dimethyl formamide as a solubilizing agent.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Food Science & Technology (AREA)
- Diabetes (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19691943580 DE1943580B2 (de) | 1969-08-27 | 1969-08-27 | Reagens zur blutzuckerbestimmung |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US3653836A true US3653836A (en) | 1972-04-04 |
Family
ID=5743926
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US66953A Expired - Lifetime US3653836A (en) | 1969-08-27 | 1970-08-26 | Reagent for determining blood sugar |
Country Status (11)
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3853469A (en) * | 1972-08-30 | 1974-12-10 | Medico Electronic Inc | Reagent and method for phosphorus determination |
| US3853473A (en) * | 1972-08-30 | 1974-12-10 | Medico Electronic Inc | Reagent and method for urea determination |
| US4425427A (en) | 1980-03-13 | 1984-01-10 | Vitafin N.V. | Simultaneous, kinetic, spectrophotometric analysis of blood serum for multiple components |
| WO1986003585A1 (en) * | 1984-12-11 | 1986-06-19 | Lawrence Paul J | Fecal occult blood test reagents and methods |
| US4818702A (en) * | 1986-06-02 | 1989-04-04 | Litmus Concepts, Inc. | Fecal occult blood test reagent |
| US5426032A (en) * | 1986-08-13 | 1995-06-20 | Lifescan, Inc. | No-wipe whole blood glucose test strip |
| US6458326B1 (en) | 1999-11-24 | 2002-10-01 | Home Diagnostics, Inc. | Protective test strip platform |
| US6525330B2 (en) | 2001-02-28 | 2003-02-25 | Home Diagnostics, Inc. | Method of strip insertion detection |
| US6541266B2 (en) | 2001-02-28 | 2003-04-01 | Home Diagnostics, Inc. | Method for determining concentration of an analyte in a test strip |
| US6562625B2 (en) | 2001-02-28 | 2003-05-13 | Home Diagnostics, Inc. | Distinguishing test types through spectral analysis |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1098808A (en) * | 1977-03-14 | 1981-04-07 | Charles T. W. Lam | Test composition and device for determining peroxidatively active substances |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3453180A (en) * | 1965-08-02 | 1969-07-01 | Miles Lab | Test article |
-
1969
- 1969-08-27 DE DE19691943580 patent/DE1943580B2/de active Pending
-
1970
- 1970-06-22 IL IL34773A patent/IL34773A/en unknown
- 1970-07-08 BR BR220400/70A patent/BR7020400D0/pt unknown
- 1970-07-27 ZA ZA705187*A patent/ZA705187B/xx unknown
- 1970-08-03 NL NL7011423A patent/NL7011423A/xx unknown
- 1970-08-25 GB GB40791/70A patent/GB1261477A/en not_active Expired
- 1970-08-25 CH CH1266870A patent/CH538118A/de not_active IP Right Cessation
- 1970-08-26 US US66953A patent/US3653836A/en not_active Expired - Lifetime
- 1970-08-26 SE SE11574/70A patent/SE360176B/xx unknown
- 1970-08-27 JP JP45075242A patent/JPS494356B1/ja active Pending
- 1970-08-27 FR FR7031383A patent/FR2059346A5/fr not_active Expired
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3453180A (en) * | 1965-08-02 | 1969-07-01 | Miles Lab | Test article |
Non-Patent Citations (5)
| Title |
|---|
| Chem. Abs. 67: 50965y (1967). * |
| Chem. Abs. 69: 103698n (1968). * |
| Chem. Abs. 69: 93549y (1968). * |
| Chem. Abs. 71: 10133a (1969). * |
| Chem. Abs. 71: 27817z (1969). * |
Cited By (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3853469A (en) * | 1972-08-30 | 1974-12-10 | Medico Electronic Inc | Reagent and method for phosphorus determination |
| US3853473A (en) * | 1972-08-30 | 1974-12-10 | Medico Electronic Inc | Reagent and method for urea determination |
| US4425427A (en) | 1980-03-13 | 1984-01-10 | Vitafin N.V. | Simultaneous, kinetic, spectrophotometric analysis of blood serum for multiple components |
| WO1986003585A1 (en) * | 1984-12-11 | 1986-06-19 | Lawrence Paul J | Fecal occult blood test reagents and methods |
| US4818702A (en) * | 1986-06-02 | 1989-04-04 | Litmus Concepts, Inc. | Fecal occult blood test reagent |
| US20030073152A1 (en) * | 1986-08-13 | 2003-04-17 | Roger Phillips | Minimum procedure system for the determination of analytes |
| US6858401B2 (en) | 1986-08-13 | 2005-02-22 | Lifescan, Inc. | Minimum procedure system for the determination of analytes |
| US5843692A (en) * | 1986-08-13 | 1998-12-01 | Lifescan, Inc. | Automatic initiation of a time interval for measuring glucose concentration in a sample of whole blood |
| US6268162B1 (en) | 1986-08-13 | 2001-07-31 | Lifescan, Inc. | Reflectance measurement of analyte concentration with automatic initiation of timing |
| US6887426B2 (en) | 1986-08-13 | 2005-05-03 | Roger Phillips | Reagents test strip adapted for receiving an unmeasured sample while in use in an apparatus |
| US6881550B2 (en) | 1986-08-13 | 2005-04-19 | Roger Phillips | Method for the determination of glucose employing an apparatus emplaced matrix |
| US20030054427A1 (en) * | 1986-08-13 | 2003-03-20 | Roger Phillips | Minimum procedure system for the determination of analytes |
| US5563042A (en) * | 1986-08-13 | 1996-10-08 | Lifescan, Inc. | Whole blood glucose test strip |
| US20030073151A1 (en) * | 1986-08-13 | 2003-04-17 | Roger Phillips | Minimum procedure system |
| US20030073153A1 (en) * | 1986-08-13 | 2003-04-17 | Roger Phillips | Minimum procedure system for the determination of analytes |
| US5426032A (en) * | 1986-08-13 | 1995-06-20 | Lifescan, Inc. | No-wipe whole blood glucose test strip |
| US6821483B2 (en) | 1986-08-13 | 2004-11-23 | Lifescan, Inc. | Reagents test strip with alignment notch |
| US6458326B1 (en) | 1999-11-24 | 2002-10-01 | Home Diagnostics, Inc. | Protective test strip platform |
| US6979571B2 (en) | 1999-11-24 | 2005-12-27 | Home Diagnostics, Inc. | Method of using a protective test strip platform for optical meter apparatus |
| US6562625B2 (en) | 2001-02-28 | 2003-05-13 | Home Diagnostics, Inc. | Distinguishing test types through spectral analysis |
| US6541266B2 (en) | 2001-02-28 | 2003-04-01 | Home Diagnostics, Inc. | Method for determining concentration of an analyte in a test strip |
| US6525330B2 (en) | 2001-02-28 | 2003-02-25 | Home Diagnostics, Inc. | Method of strip insertion detection |
| US7390665B2 (en) | 2001-02-28 | 2008-06-24 | Gilmour Steven B | Distinguishing test types through spectral analysis |
Also Published As
| Publication number | Publication date |
|---|---|
| CH538118A (de) | 1973-06-15 |
| NL7011423A (enrdf_load_stackoverflow) | 1971-03-02 |
| SE360176B (enrdf_load_stackoverflow) | 1973-09-17 |
| ZA705187B (en) | 1971-05-27 |
| DE1943580B2 (de) | 1971-08-05 |
| FR2059346A5 (enrdf_load_stackoverflow) | 1971-05-28 |
| GB1261477A (en) | 1972-01-26 |
| DE1943580A1 (de) | 1971-04-08 |
| IL34773A0 (en) | 1970-08-19 |
| IL34773A (en) | 1973-05-31 |
| BR7020400D0 (pt) | 1973-03-20 |
| JPS494356B1 (enrdf_load_stackoverflow) | 1974-01-31 |
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