SU797592A3 - Способ определени субстрата или фер-МЕНТАТиВНОй АКТиВНОСТи B биОлОгичЕСКОММАТЕРиАлЕ - Google Patents
Способ определени субстрата или фер-МЕНТАТиВНОй АКТиВНОСТи B биОлОгичЕСКОММАТЕРиАлЕ Download PDFInfo
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- SU797592A3 SU797592A3 SU772481956A SU2481956A SU797592A3 SU 797592 A3 SU797592 A3 SU 797592A3 SU 772481956 A SU772481956 A SU 772481956A SU 2481956 A SU2481956 A SU 2481956A SU 797592 A3 SU797592 A3 SU 797592A3
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- ascorbate
- ascorbate oxidase
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- 239000000758 substrate Substances 0.000 title claims abstract description 5
- 238000000034 method Methods 0.000 title claims description 15
- 239000012620 biological material Substances 0.000 title claims description 4
- 230000000694 effects Effects 0.000 title description 2
- 108010024957 Ascorbate Oxidase Proteins 0.000 claims abstract description 21
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- 238000006479 redox reaction Methods 0.000 claims abstract description 4
- 102000004190 Enzymes Human genes 0.000 claims abstract description 3
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- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 3
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 2
- 241000219130 Cucurbita pepo subsp. pepo Species 0.000 claims description 2
- 235000003954 Cucurbita pepo var melopepo Nutrition 0.000 claims description 2
- 229910052802 copper Inorganic materials 0.000 claims description 2
- 239000010949 copper Substances 0.000 claims description 2
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- 230000015572 biosynthetic process Effects 0.000 claims 1
- 125000003831 tetrazolyl group Chemical group 0.000 claims 1
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 abstract description 31
- 235000010323 ascorbic acid Nutrition 0.000 abstract description 15
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- 238000005259 measurement Methods 0.000 abstract description 6
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- JGBAASVQPMTVHO-UHFFFAOYSA-N 2,5-dihydroperoxy-2,5-dimethylhexane Chemical compound OOC(C)(C)CCC(C)(C)OO JGBAASVQPMTVHO-UHFFFAOYSA-N 0.000 abstract 1
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- 210000003608 fece Anatomy 0.000 abstract 1
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- 239000000654 additive Substances 0.000 description 2
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- IEQAICDLOKRSRL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-dodecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO IEQAICDLOKRSRL-UHFFFAOYSA-N 0.000 description 1
- JRBJSXQPQWSCCF-UHFFFAOYSA-N 3,3'-Dimethoxybenzidine Chemical compound C1=C(N)C(OC)=CC(C=2C=C(OC)C(N)=CC=2)=C1 JRBJSXQPQWSCCF-UHFFFAOYSA-N 0.000 description 1
- PCDWFBFHIIKIPM-UHFFFAOYSA-N 3-ethyl-2h-1,3-benzothiazole-2-sulfonic acid Chemical compound C1=CC=C2N(CC)C(S(O)(=O)=O)SC2=C1 PCDWFBFHIIKIPM-UHFFFAOYSA-N 0.000 description 1
- POJWUDADGALRAB-PVQJCKRUSA-N Allantoin Natural products NC(=O)N[C@@H]1NC(=O)NC1=O POJWUDADGALRAB-PVQJCKRUSA-N 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
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- 229960000458 allantoin Drugs 0.000 description 1
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- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
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- 238000012353 t test Methods 0.000 description 1
- -1 tetrazole salt Chemical class 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/54—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving glucose or galactose
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
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Description
(54) СПОСОБ ОПРЕДЕЛЕНИЯ СУБСТРАТА ИЛИ ФЕРМЕНТАТИВНОЙ АКТИВНОСТИ В БИОЛОГИЧЕСКОМ МАТЕРИАЛЕ ЧТО 0,00573 или 0,000573% аскорбата практически полностью тормоз т тест до 41,5% кюветы 4 и 5-пок зывают полное устранение нарушени этих концентраций аскорбата добавкой по 0,0003% аскорбатоксидазы. П р и м е р 2. Обнаружение глюко с 2,2,-азино-ди(3-этил-бензтиазолин сульфонатомСб) ) iftSTS), РОТЭ и got) в фотометре, с длиной волны 432 им, с температурой измерени 25°С (см. табл. 2) . Кювета i соответствует ненарушен ному измерению (нет аскорбата) .Кюве ты 2 и 3 показывают, что 0,000573% или 0,0000573. мол.% аскорбата полностью тормоз т тест до 2,1%. Кюветы 4 и 5 показывают полное устранение нарушени этих концентра ций аскорбата добавкой по 0,0003% аскорбатоксидазы. Примерз. Обнаружение мочевой кислоты посредством фенола, ами ноантипирина, POD и урикаэы на аналитическом автомате (Auto Analyzar) Принцип тестам мочева кислота + -J- Оа + HaQ Р аллантоин + + HjOj - 2 H2.Og + 2,4-дихлорфенал + 4-аминоантипирин хинодный кр ситель + 2 HgO . Изготовление растворов. Реактив аскорба оксидаэы. В 600 бидистилдированной воды раствор етс содержимое флакона 1. Добавка 0,3 м Brij-35. Затем смесь вытверживаетс в темном флаконе приблизительно при четыре недели,при одну неделю . Реактив уриказы. В 800 мл бидистиллированной воды раствор етс содержимое флакона 2. Добавка 2,0 мл В г Ij-35 . Затем смесь выдерживаетс в темном флаконе приблизительно при 4с четыре недели, приблизительно при 25°С одну неделю. Концентраци растворов. 0,63% фосфатного буферного раствора , рН 5,6 . 0,35% трис/ликюнной кислоты, рН 8,9, уриказы 0,001%, POD 0,000015%, 0,05% 2,4-дихлорфенола , 0,08% 4-аминоантипирина. П р и м е р 4. Обнаружение глюкозы с HK/Q 6Р-ОА, NADP, INT и диафоразой в фортометре. Измер ема длина волны 492 нм, температура инкубации 25С (см. табл. 3). Кювета 1 соответствует ненарушенному измерению. Кювета 2 показывает , что 0,0000573% аскорбата может тормозить тест на 34% кювета 3 показывает , что 0,0003% аскорбатоксидазы полностью устран ет это нарушение . П р и м е р 5. Определение глутамата посредством диафоразы в фотометре . Измер ема длина волны 492 нм, температура 25°С(см. табл. 4}, Кювета 1 соответствует ненарушенному измерению. кюветы 2 и 3 показывают, что 0,0029% или 0,00029% аскорбата тормоз т тест на 700% или на 100%. Кюветы 4 и 5 показывают полное устранение этого нарушени добавкой 0,00006% аскорбатоксидазы. Примере. Определение тирозина с 3-метил-б калийсульфонил-бензтиазолон-гидра-зоном- (2) (SMBTH) и тирозиназой в фотометре, измер ема длина волны 492 нм, измер ема температура 25Ос(см. табл. 5. Кювета 1 соответствует ненарушенному измерению, в кювете 2 0,0057% аскорбата снижает теоретическую величану на 35%, в кювете 3 это нарушение полностью устран етс добавкой 0,003% аскорбатоксидазы. П р и м.е р 7. Определение пирокатехина с SMBTH и дифенол-оксидаЗОЙ (см. табл. 6) . Кювета 1 соответствует ненарушенному измерению. В кювете 20,01145% аскорбата снижают теоретическую величину на 47% это нарушение полностью устран етс в кювете 3 добавкой 0,0003% аскорбатоксидаэы . Полученный способ позвол ет полностью устранить вызванные ошибки наличием аскорбиновой кислоты при проведении окислительно-восстановительных реакций. Таблица 1
Фосфатный буферный раствор, рН 7,01, 1,368%
о-Дианизидин/, 0,5% POD, 0,18%
Раствор глюкозы, 0,172%
Раствор аскорбиновой кислоты
Вода
Аскорбатоксидаза 500 ед./мл, 0,05%
Инкубаци 1 мин при , считывание Е , затем начинаетс реакци с
GOD 0,0333% Инкубаци 30 мин при &Е Инкубаци 30 мин при считывание Е,, дЕ0,505 0,000
Продолжение табл. 1
0,03 0,03 0,030,030,03
0,1 0,010,10,01
0,12 0,02 0,11 -0,09
-0,02. 0,02
0,1 0,1 0,1 0,1 0,1 ,считывание Е,вычисление из & 0,541 0,010 0,316 0,540 0,543 вычисление из АЕ 0,400,502 0,504
Фосфатный буферный раствор, рН 7,5, 1,36%
NADP INT по 0,1% Диафораза 5 Е/мп, 0,1% Раствор глюкозы, 0,005% Раствор аскорбата 0,17%
Аскорбатоксидаза, 0,00035%
Вода
Инкубаци 3 мин при 25с, считывание Е на 1ало реакции с HK/G6P-DH-pacTBop по 0,14%0,050,05 0,05
Инкубаци 15 мин, считывание Е, вычисление из Ej-E ДЕ ДЕ0,2340,307 0,230
Т а ti ) и
0,03
0,04
0,03
Таблица 4 Считывание IT- раствор 0,2 Инкубаци 15 мин при 25с, считывание
ЛЕ
Компоненты
Фосфатный буферный, раствор рН 5,2, 3,4%
SMBTH 0,1 мол, 3%
Раствор аскорбиновой кислоты. О,112%
Вода
Аскорбатоксидаза 500 ,05%
Тирозиназа, 0,2 Инкубаци 1 мин при 0,036% тирозин Инкубаци 1 час при ДЕ
Продолжение табл. 4
0,184 1,560 0,380 0,186 0,182
iT а б л и ц а 5
Юовета
2,77
2,77 0,05 0,05
0/1
0,1 0,02
у
0,02 0,05
0,05 Е , начало реакции с 0,05 0,05 0,05 0,05 E/j вычисление из АЕ считываниеЕ , начало реакции с 0,050,05 0,05 считываниеЕ, вычисление из Е2-Е ЛЕ 1,1130,728 1,100
Компоненты Инкубаци 1 мин при , считывание дефенолоксидаза, 0,2% 0,05 Инкубаци 17 мин при 25-с считывание ЛЕ0,890 формула изобретени 1.Способ определени субстрата или ферментативной активности в био логическом материале путем проведени окислительно-восстановительной реакции, отличающийс тем,что, с целью повьацени точности способа, в реакционную смесь внос т аскбрбатоксйдазу в количестве 100 Eff/мп исходной смеси, а учет ре акции ведут по образованию окрашенного продукта. 2.Способ по п.1, отличающийс тем, что дл получени окрашенного продукта аскорбатоксидазу ввод т одновременно с Н О образукхцим ферментом.
Продолжение табл..
Кювета Е , начало реакции с 0,05 0,05 Е вычисление из Е„-Е. ДЕ 0,485 Г,896 3. Способ по п.1, отличающийс тем, что дл получени окрашенного продукта аскорбатоксидазу ввод т одновременно с солью тетразоли . 4. Способ по П.1, отлича ющ и и с т-эм, что дл получени окрашенного продукта аскорбатокси дазу ввод т одновременно с фенолом. 5, Способ по П.1, отличающийс тем, что используют аскорбатоксидазу из Zucchini (Cucurbuta реро medunos,a). Источники информации, йрин тые во внимание при экспертизе 1. Biochemical Copper Proceedings. «tarriman, N.V. 1965, с. 305-337.
Claims (5)
- формула изобретения1. Способ определения субстрата или ферментативной активности в биологическом материале путем проведения окислительно-восстановительной реакции, отличающийся тем,что, с целью повышения точности способа, в реакционную смесь вносят аскорбатоксидазу в количестве100 Е«/мл исходной смеси, а учет реакцшгведут по образованию окрашенного продукта.
- 2. Способ по п.1, отличающийся тем, что для получения окрашенного продукта аскорбатоксидазу вводят одновременно с Н^О^образукщим ферментом.
- 3. Способ по п.1, отличающийся тем, что для получения окрашенного продукта аскорбатоксидазу вводят одновременно с солью тетразолия.
- 4. Способ по п.1, отличающийся тем, что для получения окрашенного продукта аскорбатоксидаэу вводят одновременно с фенолом.
- 5. Способ по п.1, отличающий с я тем, что используют аскорбатоксидазу из Zucchini (Cucurbuta pepo medu J1oia).30 Источники информации,Принятые во внимание при экспертизе1. Biochemical Copper Proceedings.Harriman, N.V. 1965, c. 305-337.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2625834A DE2625834B2 (de) | 1976-06-09 | 1976-06-09 | Verfahren zur Bestimmung von Substraten oder Enzymaktivitäten |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SU797592A3 true SU797592A3 (ru) | 1981-01-15 |
Family
ID=5980154
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SU772481956A SU797592A3 (ru) | 1976-06-09 | 1977-05-06 | Способ определени субстрата или фер-МЕНТАТиВНОй АКТиВНОСТи B биОлОгичЕСКОММАТЕРиАлЕ |
Country Status (29)
| Country | Link |
|---|---|
| US (1) | US4168205A (ru) |
| JP (1) | JPS52150692A (ru) |
| AR (1) | AR212763A1 (ru) |
| AT (1) | AT354640B (ru) |
| AU (1) | AU500988B2 (ru) |
| BE (1) | BE854407A (ru) |
| BR (1) | BR7703008A (ru) |
| CA (1) | CA1084393A (ru) |
| CH (1) | CH633887A5 (ru) |
| CS (2) | CS235068B2 (ru) |
| DD (1) | DD130178A5 (ru) |
| DE (1) | DE2625834B2 (ru) |
| DK (1) | DK144949C (ru) |
| ES (1) | ES458544A1 (ru) |
| FI (1) | FI60719C (ru) |
| FR (1) | FR2354561A1 (ru) |
| GB (1) | GB1519134A (ru) |
| HU (1) | HU172931B (ru) |
| IE (1) | IE45546B1 (ru) |
| IL (1) | IL52047A (ru) |
| IT (1) | IT1075527B (ru) |
| LU (1) | LU77290A1 (ru) |
| NL (1) | NL169503C (ru) |
| NO (1) | NO148340C (ru) |
| PL (2) | PL109224B1 (ru) |
| SE (1) | SE426601B (ru) |
| SU (1) | SU797592A3 (ru) |
| YU (1) | YU114477A (ru) |
| ZA (1) | ZA772716B (ru) |
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| DE2914487A1 (de) * | 1979-04-10 | 1980-10-30 | Boehringer Mannheim Gmbh | Verfahren und mittel zur entfernung von ascorbinsaeure aus waessrigen fluessigkeiten |
| JPS56109595A (en) * | 1980-02-01 | 1981-08-31 | Wako Pure Chem Ind Ltd | Removal of reductive inhibitor |
| DE3012368C2 (de) * | 1980-03-29 | 1982-04-15 | Boehringer Mannheim Gmbh, 6800 Mannheim | Verfahren und diagnostische Mittel zum Nachweis von Redox-Reaktionen |
| DE3012314A1 (de) * | 1980-03-29 | 1981-10-15 | W. Schlafhorst & Co, 4050 Mönchengladbach | Offenend-spinnvorrichtung |
| US4310626A (en) * | 1980-06-02 | 1982-01-12 | Miles Laboratories, Inc. | Interference-resistant composition, device and method for determining a peroxidatively active substance in a test sample |
| DE3046741A1 (de) * | 1980-12-11 | 1982-07-15 | Boehringer Mannheim Gmbh, 6800 Mannheim | Nachweis von nad(p)h oder salicylat |
| DE3249743C2 (ru) * | 1982-02-18 | 1990-10-04 | Amano Pharmaceutical Co., Ltd., Nagoya, Aichi, Jp | |
| US4587220A (en) * | 1983-03-28 | 1986-05-06 | Miles Laboratories, Inc. | Ascorbate interference-resistant composition, device and method for the determination of peroxidatively active substances |
| DE3313178A1 (de) * | 1983-04-12 | 1984-10-18 | Boehringer Mannheim Gmbh, 6800 Mannheim | Verfahren zur enzymatischen bestimmung von sulfit |
| JPS59230161A (ja) * | 1983-06-13 | 1984-12-24 | Wako Pure Chem Ind Ltd | 還元性物質の分解方法及び分解用試薬 |
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| US5196314A (en) * | 1986-04-04 | 1993-03-23 | Boehringer Mannheim Gmbh | Process and reagent for the determination of substrates or enzyme activities |
| DE3620817A1 (de) * | 1986-06-21 | 1987-12-23 | Boehringer Mannheim Gmbh | Verfahren zur spezifischen bestimmung des serumfructosamingehalts sowie hierfuer geeignetes reagenzgemisch |
| US4885240A (en) * | 1986-09-04 | 1989-12-05 | Eastman Kodak Company | Use of organic buffers to reduce dehydroascorbic acid interference in analytical methods |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3411887A (en) * | 1964-06-15 | 1968-11-19 | Miles Lab | Diagnostic composition |
| AT308049B (de) * | 1970-08-28 | 1973-06-25 | Boehringer Mannheim Gmbh | Reagens zur Glucosebestimmung |
| JPS5033795B1 (ru) * | 1970-11-25 | 1975-11-04 | ||
| US3791988A (en) * | 1972-03-23 | 1974-02-12 | Hoffmann La Roche | Diagnostic test for glucose |
| US3992158A (en) * | 1973-08-16 | 1976-11-16 | Eastman Kodak Company | Integral analytical element |
| FR2280081A1 (fr) * | 1974-07-23 | 1976-02-20 | Kodak Pathe | Produit composite unitaire pour l'analyse chimique ou biologique |
-
1976
- 1976-06-09 DE DE2625834A patent/DE2625834B2/de active Granted
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1977
- 1977-04-04 AT AT234077A patent/AT354640B/de not_active IP Right Cessation
- 1977-04-13 IE IE764/77A patent/IE45546B1/en not_active IP Right Cessation
- 1977-04-13 CA CA276,113A patent/CA1084393A/en not_active Expired
- 1977-04-25 AR AR267337A patent/AR212763A1/es active
- 1977-04-27 NL NLAANVRAGE7704591,A patent/NL169503C/xx not_active IP Right Cessation
- 1977-04-28 US US05/792,073 patent/US4168205A/en not_active Expired - Lifetime
- 1977-04-28 FI FI771356A patent/FI60719C/fi not_active IP Right Cessation
- 1977-04-29 GB GB17993/77A patent/GB1519134A/en not_active Expired
- 1977-05-03 IT IT77@@A patent/IT1075527B/it active
- 1977-05-05 YU YU01144/77A patent/YU114477A/xx unknown
- 1977-05-06 HU HU77BO00001667A patent/HU172931B/hu unknown
- 1977-05-06 CS CS781874A patent/CS235068B2/cs unknown
- 1977-05-06 JP JP5195377A patent/JPS52150692A/ja active Granted
- 1977-05-06 SU SU772481956A patent/SU797592A3/ru active
- 1977-05-06 PL PL1977197927A patent/PL109224B1/pl unknown
- 1977-05-06 IL IL52047A patent/IL52047A/xx unknown
- 1977-05-06 ES ES458544A patent/ES458544A1/es not_active Expired
- 1977-05-06 CH CH571277A patent/CH633887A5/de not_active IP Right Cessation
- 1977-05-06 SE SE7705303A patent/SE426601B/xx not_active IP Right Cessation
- 1977-05-06 PL PL1977217830A patent/PL114343B1/pl unknown
- 1977-05-06 NO NO771599A patent/NO148340C/no unknown
- 1977-05-06 CS CS773018A patent/CS207453B2/cs unknown
- 1977-05-06 ZA ZA00772716A patent/ZA772716B/xx unknown
- 1977-05-09 LU LU77290A patent/LU77290A1/xx unknown
- 1977-05-09 FR FR7714090A patent/FR2354561A1/fr active Granted
- 1977-05-09 AU AU25021/77A patent/AU500988B2/en not_active Expired
- 1977-05-09 BE BE177392A patent/BE854407A/xx not_active IP Right Cessation
- 1977-05-09 BR BR7703008A patent/BR7703008A/pt unknown
- 1977-05-09 DK DK202677A patent/DK144949C/da not_active IP Right Cessation
- 1977-05-09 DD DD7700198825A patent/DD130178A5/xx unknown
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