US3716632A - Test composition and method for the in vitro determination of thyroid function - Google Patents
Test composition and method for the in vitro determination of thyroid function Download PDFInfo
- Publication number
- US3716632A US3716632A US00032396A US3716632DA US3716632A US 3716632 A US3716632 A US 3716632A US 00032396 A US00032396 A US 00032396A US 3716632D A US3716632D A US 3716632DA US 3716632 A US3716632 A US 3716632A
- Authority
- US
- United States
- Prior art keywords
- indicator
- composition
- blue
- thyroid
- test 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
- 238000012360 testing method Methods 0.000 title claims abstract description 28
- 210000001685 thyroid gland Anatomy 0.000 title claims abstract description 15
- 239000000203 mixture Substances 0.000 title claims description 31
- 238000000034 method Methods 0.000 title claims description 21
- 238000000338 in vitro Methods 0.000 title abstract description 4
- 230000002285 radioactive effect Effects 0.000 claims abstract description 21
- AUYYCJSJGJYCDS-LBPRGKRZSA-N Thyrolar Chemical class IC1=CC(C[C@H](N)C(O)=O)=CC(I)=C1OC1=CC=C(O)C(I)=C1 AUYYCJSJGJYCDS-LBPRGKRZSA-N 0.000 claims abstract description 18
- ABIUHPWEYMSGSR-UHFFFAOYSA-N bromocresol purple Chemical compound BrC1=C(O)C(C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C=C(Br)C(O)=C(C)C=2)=C1 ABIUHPWEYMSGSR-UHFFFAOYSA-N 0.000 claims abstract description 17
- 210000002966 serum Anatomy 0.000 claims abstract description 16
- 239000007793 ph indicator Substances 0.000 claims abstract description 15
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 12
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 12
- 238000001179 sorption measurement Methods 0.000 claims abstract description 10
- 239000005495 thyroid hormone Substances 0.000 claims abstract description 8
- 229940036555 thyroid hormone Drugs 0.000 claims abstract description 8
- 239000000126 substance Substances 0.000 claims abstract description 5
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 claims description 15
- NUHCTOLBWMJMLX-UHFFFAOYSA-N bromothymol blue Chemical compound BrC1=C(O)C(C(C)C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C(=C(Br)C(O)=C(C(C)C)C=2)C)=C1C NUHCTOLBWMJMLX-UHFFFAOYSA-N 0.000 claims description 13
- 229940035722 triiodothyronine Drugs 0.000 claims description 9
- FRPHFZCDPYBUAU-UHFFFAOYSA-N Bromocresolgreen Chemical compound CC1=C(Br)C(O)=C(Br)C=C1C1(C=2C(=C(Br)C(O)=C(Br)C=2)C)C2=CC=CC=C2S(=O)(=O)O1 FRPHFZCDPYBUAU-UHFFFAOYSA-N 0.000 claims description 8
- 239000000872 buffer Substances 0.000 claims description 8
- WWAABJGNHFGXSJ-UHFFFAOYSA-N chlorophenol red Chemical compound C1=C(Cl)C(O)=CC=C1C1(C=2C=C(Cl)C(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 WWAABJGNHFGXSJ-UHFFFAOYSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 7
- 230000002401 inhibitory effect Effects 0.000 claims description 5
- 239000004698 Polyethylene Substances 0.000 claims description 4
- -1 polyethylene Polymers 0.000 claims description 4
- 229920000573 polyethylene Polymers 0.000 claims description 4
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 abstract description 11
- 238000009601 thyroid function test Methods 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 230000007423 decrease Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 13
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 239000011347 resin Substances 0.000 description 5
- 229920005989 resin Polymers 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 239000013068 control sample Substances 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- 229920002307 Dextran Polymers 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 229920005654 Sephadex Polymers 0.000 description 3
- 239000012507 Sephadex™ Substances 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 239000012085 test solution Substances 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- XUIIKFGFIJCVMT-GFCCVEGCSA-N D-thyroxine Chemical compound IC1=CC(C[C@@H](N)C(O)=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-GFCCVEGCSA-N 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 229940034208 thyroxine Drugs 0.000 description 2
- XUIIKFGFIJCVMT-UHFFFAOYSA-N thyroxine-binding globulin Natural products IC1=CC(CC([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-UHFFFAOYSA-N 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 206010020850 Hyperthyroidism Diseases 0.000 description 1
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 1
- 102000009488 Thyroxine-Binding Proteins Human genes 0.000 description 1
- 108010048889 Thyroxine-Binding Proteins Proteins 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002085 enols Chemical class 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 230000005251 gamma ray Effects 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 208000003532 hypothyroidism Diseases 0.000 description 1
- 230000002989 hypothyroidism Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 230000005865 ionizing radiation Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229960003531 phenolsulfonphthalein Drugs 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/78—Thyroid gland hormones, e.g. T3, T4, TBH, TBG or their receptors
-
- 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
- Y10S436/804—Radioisotope, e.g. radioimmunoassay
-
- 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
- Y10S436/826—Additives, e.g. buffers, diluents, preservatives
Definitions
- thyroxine-binding proteins are the primary binding sites for the hormones produced by the thyroid gland. Since the TBF have a strong affinity for thyroid hormone, they will bind any free thyroid hormone available until they become saturated. In this test, such hormone is made available to the TB? in a sample of the-patients serum and a secondary binding site consisting of a resin powder or crosslinked dextran (Sephadex) is provided to take up any of the added radioactive triiodothyronine or thyroxine (hereinafter referred to as T or T respectively) not bound by the TBP.
- T or T radioactive triiodothyronine
- the main object of the present invention is to provide a thyroid function test solution containing a pH indicator which possesses the three fold advantage of (1) providing a color change when the serum sample is added thereto (2) inhibiting the formation of free iodide from radioactive iodinated T and (3) decreasing the adsorption of radioactive iodinated T to the walls of the vessel containing the same.
- thyroid function tests may be performed in many different ways, one common method involves adding an appropriate amount ofa buffered aqueous solu tion of radioactive triiodothyronine or thyroxine and serum to a column of ion exchange resin such as Amberlite IRA 400 made by Rohm and Haas Co. of Philadelphia, Pa. or a cross linked dextran such as Sephadex G-25 made by Pharmacia Corp. of Uppsala, Sweden.
- the column is then drained and the radioactivity measured in a gamma ray counting instrument for 1 minute.
- the column is thereafter eluted with distilled water and the radioactivity remaining on the column is again measured for 1 minute.
- the percent retention is then calculated by comparing the two readings. Retentions of 41 percent to 6l percent are considered normal with cross-linked dextran (Sephadex) columns and 25 percent to 35 percent with resin columns.
- the pH indicator employed in the present invention is of the protein error type which changes color in the presence of albumin without a significant actual change in pH.
- an indicator is added in a concentration of about 5. to micrograms per milliliter of aqueous I -T reagent buffered at a pH of 5 the reagent changes from its initial color to a second color when the serum is added.
- This second color acts as a marker which will appear at varying heights on the resin or dextrancolumndepending on the last operation step completed in the test procedure, since the color will move with the solvent front.
- a preferred indicator is bromcresol purple which changes color from yellow to purple within the pH range of 5.2 to 6.8.
- the yellow-orange color will turn to a grey-green color when serum is added at a ratio of 0.05 milliliter of serum to 0.45 milliliter of reagent mixture.
- the green color indicates the presence of the serum.
- the improved T test reagent with pH indicator of this invention will contain the following:
- a buffered aqueous solution of T containing 60 micrograms of bromcresol purple per milliliter prepared as shown in the foregoing example was stored under various conditions in a polyethylene container and the percent of free iodide formed was compared with a similar solution containing no pH indicator.
- the initial solutions employed contained about 2 percent free iodide.
- the radioactive T solution containing the indicator showed 2.25 percent free iodide compared to 5.2 percent free iodide in the control sample containing no indicator.
- the T test solution with pH indicator contained 4.25 percent free iodide, whereas the control sample had a free iodide content of 9.0 percent.
- a still further advantage of the use of a pH indicator in T test solution is to inhibit the adsorption of radioactive T to the walls of a plastic or glass container.
- Propylene glycol and albumin have been used in the past to reduce such adsorption but these additives interfere with the thyroid function test.
- aqueous radioactive T solutions containing one of several indicators at a concentration of 60 micrograms per milliliter of solution were rotated on a wheel in a polyethylene container for 14 days and the percent activity remaining was compared with a control sample containing no indicator.
- prewashed means contacting (such as by rinsing, soaking or flushing) the interior surfaces of said containers with an aqueous, buffered solution of one of the pH indicators hereinbefore disclosed.
- Bromcresol purple or bromthymol blue are the preferred indicators. These are employed at a concentration of from 50 to 70 micrograms per milliliter of water buffered to a pH of about 5.
- T employed in the test compositions of this invention can be made radioactive with either F or 1", although l is preferred, since it has a half-life of 56 days compared to 1"" which has a half-life of 8 days.
- a test composition for determining thyroid function comprising a radioactive thyroid hormone and a buffer for establishing the pH of thecomposition in a predetermined range
- the improvement which comprises adding to the composition a color responsive is used to maintain the pH of the composition at about 5 3.
- the indicator is selected from the group consistin of bromcresol purple, chlorphenol red, tetrabromp enol blue,
- bromphenol blue bromcresol green and bromthymol blue.
- test composition as in claim 1 wherein the indicator is bromcresol purple.
- a test composition as in claim 1 wherein the indicator is bromthymol blue.
- a test composition as in claim 1 wherein the thyroid substance is triiodothyronine.
- a method as in claim 8 wherein the indicator is selected from the group consisting of bromcresol purple, chlorphenol red, tetrabromphenol blue, bromphenol blue, bromcresol green and bromthymol blue.
- a method as in claim 8 wherein the thyroid substance is triiodothyronine.
- a method of inhibiting the adsorption of radioactive triiodothyronine from an aqueous solution thereof on the walls of a polyethylene container which comprises adding to said aqueous solution an adsorption inhibiting amount of a protein error type pH indicator.
- a method as in claim 13 wherein the indicator is selected from the group consisting of bromcresol purple, chlorphenol red, tetrabromphenol blue, bromphenol blue, bromcresol green and bromthymol blue.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (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 |
|---|---|---|---|
| US3239670A | 1970-04-27 | 1970-04-27 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US3716632A true US3716632A (en) | 1973-02-13 |
Family
ID=21864745
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US00032396A Expired - Lifetime US3716632A (en) | 1970-04-27 | 1970-04-27 | Test composition and method for the in vitro determination of thyroid function |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US3716632A (OSRAM) |
| CA (1) | CA956097A (OSRAM) |
| DE (1) | DE2120442B2 (OSRAM) |
| FR (1) | FR2090714A5 (OSRAM) |
| GB (1) | GB1347498A (OSRAM) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3960492A (en) * | 1974-05-31 | 1976-06-01 | Nuclear Diagnostics, Inc. | Method for determining an index of binding protein content of blood |
| US3985867A (en) * | 1974-01-02 | 1976-10-12 | The Radiochemical Centre Limited | Immunoassays employing a colored second antibody |
| US4108974A (en) * | 1976-08-27 | 1978-08-22 | Bio-Rad Laboratories, Inc. | Radioimmunoassay for thyroid hormone |
| US4278653A (en) * | 1979-02-01 | 1981-07-14 | New England Nuclear Corporation | Methods and kits for double antibody immunoassay providing a colored pellet for easy visualization |
| US4299812A (en) * | 1978-11-29 | 1981-11-10 | Diagnostic Products Corp. | Immunoassay of thyroxine in neonates using dried blood samples |
| US4311690A (en) * | 1978-06-20 | 1982-01-19 | Damon Corporation | Test set and method for the determination of free hormones |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3485587A (en) * | 1956-02-29 | 1969-12-23 | Miles Lab | Protein indicator |
| US3507618A (en) * | 1964-11-27 | 1970-04-21 | Squibb & Sons Inc | Apparatus and method for determining thyroid function |
| US3516794A (en) * | 1964-12-14 | 1970-06-23 | Squibb & Sons Inc | Apparatus and method for determining thyroid function |
-
1970
- 1970-04-27 US US00032396A patent/US3716632A/en not_active Expired - Lifetime
-
1971
- 1971-03-24 CA CA108,577A patent/CA956097A/en not_active Expired
- 1971-04-26 FR FR7114752A patent/FR2090714A5/fr not_active Expired
- 1971-04-26 GB GB1141971*[A patent/GB1347498A/en not_active Expired
- 1971-04-26 DE DE19712120442 patent/DE2120442B2/de active Granted
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3485587A (en) * | 1956-02-29 | 1969-12-23 | Miles Lab | Protein indicator |
| US3507618A (en) * | 1964-11-27 | 1970-04-21 | Squibb & Sons Inc | Apparatus and method for determining thyroid function |
| US3516794A (en) * | 1964-12-14 | 1970-06-23 | Squibb & Sons Inc | Apparatus and method for determining thyroid function |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3985867A (en) * | 1974-01-02 | 1976-10-12 | The Radiochemical Centre Limited | Immunoassays employing a colored second antibody |
| US3960492A (en) * | 1974-05-31 | 1976-06-01 | Nuclear Diagnostics, Inc. | Method for determining an index of binding protein content of blood |
| US4108974A (en) * | 1976-08-27 | 1978-08-22 | Bio-Rad Laboratories, Inc. | Radioimmunoassay for thyroid hormone |
| US4311690A (en) * | 1978-06-20 | 1982-01-19 | Damon Corporation | Test set and method for the determination of free hormones |
| US4299812A (en) * | 1978-11-29 | 1981-11-10 | Diagnostic Products Corp. | Immunoassay of thyroxine in neonates using dried blood samples |
| US4278653A (en) * | 1979-02-01 | 1981-07-14 | New England Nuclear Corporation | Methods and kits for double antibody immunoassay providing a colored pellet for easy visualization |
Also Published As
| Publication number | Publication date |
|---|---|
| DE2120442A1 (de) | 1971-11-11 |
| GB1347498A (en) | 1974-02-27 |
| DE2120442C3 (OSRAM) | 1973-11-29 |
| FR2090714A5 (OSRAM) | 1972-01-14 |
| CA956097A (en) | 1974-10-15 |
| DE2120442B2 (de) | 1973-05-10 |
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