JPS6142826B2 - - Google Patents
Info
- Publication number
- JPS6142826B2 JPS6142826B2 JP53142815A JP14281578A JPS6142826B2 JP S6142826 B2 JPS6142826 B2 JP S6142826B2 JP 53142815 A JP53142815 A JP 53142815A JP 14281578 A JP14281578 A JP 14281578A JP S6142826 B2 JPS6142826 B2 JP S6142826B2
- Authority
- JP
- Japan
- Prior art keywords
- urea
- solution
- absorbance
- phthalaldehyde
- reagent
- 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
Links
- 238000000034 method Methods 0.000 claims description 19
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 16
- 239000004202 carbamide Substances 0.000 claims description 16
- 238000002835 absorbance Methods 0.000 claims description 11
- 239000000243 solution Substances 0.000 description 13
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- 238000010438 heat treatment Methods 0.000 description 5
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 4
- XGEGHDBEHXKFPX-UHFFFAOYSA-N N-methylthiourea Natural products CNC(N)=O XGEGHDBEHXKFPX-UHFFFAOYSA-N 0.000 description 4
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 4
- 229960002173 citrulline Drugs 0.000 description 4
- 235000013477 citrulline Nutrition 0.000 description 4
- XGEGHDBEHXKFPX-NJFSPNSNSA-N methylurea Chemical compound [14CH3]NC(N)=O XGEGHDBEHXKFPX-NJFSPNSNSA-N 0.000 description 4
- FSEUPUDHEBLWJY-HWKANZROSA-N diacetylmonoxime Chemical compound CC(=O)C(\C)=N\O FSEUPUDHEBLWJY-HWKANZROSA-N 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- LUBJCRLGQSPQNN-UHFFFAOYSA-N 1-Phenylurea Chemical compound NC(=O)NC1=CC=CC=C1 LUBJCRLGQSPQNN-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- RYECOJGRJDOGPP-UHFFFAOYSA-N Ethylurea Chemical compound CCNC(N)=O RYECOJGRJDOGPP-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 125000001637 1-naphthyl group Chemical group [H]C1=C([H])C([H])=C2C(*)=C([H])C([H])=C([H])C2=C1[H] 0.000 description 1
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N Aminoantipyrine Natural products CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 1
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 1
- 239000003929 acidic solution Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- VEQOALNAAJBPNY-UHFFFAOYSA-N antipyrine Chemical compound CN1C(C)=CC(=O)N1C1=CC=CC=C1 VEQOALNAAJBPNY-UHFFFAOYSA-N 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- SFNALCNOMXIBKG-UHFFFAOYSA-N ethylene glycol monododecyl ether Chemical compound CCCCCCCCCCCCOCCO SFNALCNOMXIBKG-UHFFFAOYSA-N 0.000 description 1
- 229960005222 phenazone Drugs 0.000 description 1
- -1 polyoxyethylene lauryl ether Polymers 0.000 description 1
- 229920000259 polyoxyethylene lauryl ether Polymers 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 239000012085 test solution Substances 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
- G01N33/62—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving urea
Description
【発明の詳細な説明】
本発明は尿素およびその誘導体の定量法に関す
る。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for quantifying urea and its derivatives.
尿素およびその誘導体に共通した比色定量法と
して実用的に用い得る方法はきわめて少く、酸性
溶液中でジアセチルモノオキシム、ジアセルモノ
オキシムとアンチピリン、またはジアセチルモノ
オキシムとチオミカルバジドにより発色させて比
色定量する方法があげられるぐらいである。しか
しながらこれらの方法はいずれも濃厚なリン酸溶
液中で100℃に加熱するか、あるいはかなり濃厚
な硫酸溶液中で37℃に長く加熱することが必要で
あつて、しかもジアセチルモノオキシムは特異な
臭気をもち、人によつては不快感を生じる。した
がつてこれらの方法は繁用に耐える良法とはいい
難い。 There are very few practical methods for colorimetric determination common to urea and its derivatives, and colorimetry is performed by developing color with diacetylmonoxime, diacylmonoxime and antipyrine, or diacetylmonoxime and thiomicarbazide in an acidic solution. I can only give you a method. However, all of these methods require heating to 100°C in a concentrated phosphoric acid solution or prolonged heating to 37°C in a fairly concentrated sulfuric acid solution, and diacetyl monooxime has a peculiar odor. and may cause discomfort to some people. Therefore, these methods cannot be said to be good methods that can withstand frequent use.
これらの点に鑑み本発明者らは種々検討の結果
尿素およびその誘導体がＮ―（１―ナフチル）―
N′―ジエチルエチレンジアミンおよび０―フタ
ルアルデヒドにより鋭敏に発色することを見出し
た。 In view of these points, the present inventors conducted various studies and found that urea and its derivatives are N-(1-naphthyl)-
It has been found that N'-diethylethylenediamine and 0-phthalaldehyde cause a sharp color development.
本発明はこれらの新知見に基づいて完成された
もので、試料にＮ―（１―ナフチル）―N′―ジ
エチルエチレンジアミンおよび０―フタルアルデ
ヒドを作用させて反応液の吸光度を測定すること
を特徴とする尿素およびその誘導体の定量法であ
る。 The present invention was completed based on these new findings, and is characterized by measuring the absorbance of the reaction solution by reacting N-(1-naphthyl)-N'-diethylethylenediamine and 0-phthalaldehyde with a sample. This is a method for quantifying urea and its derivatives.
この反応は1N塩酸中で37℃、15分間加温で行
なわれるので、操作はきわめて容易であり、試薬
取扱い上の危険も少く、臭気も発しない。反応は
短時間で終了し、反応液は赤色から橙褐色に変化
するので、反応後吸光度（465nm）を測定して比
色定量できるばかりでなく、初速度法（520nm）
を利用して定量することもできる。また試薬溶液
は安定であつて、本法は実用的な尿素およびその
誘導体の定量法として価値あるものである。 This reaction is carried out in 1N hydrochloric acid at 37°C by heating for 15 minutes, so the operation is extremely easy, there is little danger in handling reagents, and there is no odor. The reaction completes in a short time, and the reaction solution changes from red to orange-brown, so it is possible to not only perform colorimetric determination by measuring the absorbance (465 nm) after the reaction, but also use the initial velocity method (520 nm).
It can also be quantified using Furthermore, the reagent solution is stable, making this method valuable as a practical method for quantifying urea and its derivatives.
尿素の分析のために０―フタルアルデヒドとＮ
―（１―ナフチル）エチレンジアンを用いる方法
が提案されている（クリニカル・ケミストリー第
21巻、第８号、1136頁、1975年）がこの方法も多
量の濃硫酸を用いる上、ブランク値が高いことが
難点であるに比し、本発明によればこれらの難点
が一挙に解決され、上記利点を有する。 0-phthalaldehyde and N for the analysis of urea
- A method using (1-naphthyl)ethylenedian has been proposed (Clinical Chemistry Vol.
21, No. 8, p. 1136, 1975), this method also uses a large amount of concentrated sulfuric acid and has the disadvantages of high blank values, but the present invention solves these disadvantages at once. and has the above advantages.
実施例 １
試薬１の調製
Ｎ―（１―ナフチル）―N′―ジエチルエチレ
ンジアミンシユウ酸塩0.854ｇを水500mlに加温し
て溶かし、冷却後ポリオキシエチレンラウリルエ
ーテル（Brij35）１ｇを溶かす。Example 1 Preparation of Reagent 1 0.854 g of N-(1-naphthyl)-N'-diethylethylenediamine oxalate is dissolved in 500 ml of water by heating, and after cooling, 1 g of polyoxyethylene lauryl ether (Brij35) is dissolved.
試薬２の調製
０―フタルアルデヒド1.34ｇを2N塩酸500mlに
加温して溶かす。Preparation of Reagent 2 Dissolve 1.34 g of 0-phthalaldehyde in 500 ml of 2N hydrochloric acid by heating.
分析方法
尿素溶液50μに試薬１および２をそれぞれ２
mlずつ加え、37℃で15分間加温したのち流水で２
分間冷却し、空試験液を対照として波長465nmで
吸光度を測定する。この方法により尿素0.05〜
1.0μＭの範囲において尿素量と吸光度は直線関
係にあるから、予め作成した検量線から尿素量を
定量する。Analysis method: Add 2 portions each of reagents 1 and 2 to 50μ of urea solution.
Add ml at a time, warm at 37℃ for 15 minutes, and rinse with running water.
Cool for a minute and measure the absorbance at a wavelength of 465 nm using the blank test solution as a control. By this method, urea 0.05 ~
Since there is a linear relationship between the amount of urea and the absorbance in the range of 1.0 μM, the amount of urea is determined from a calibration curve prepared in advance.
フエニル尿素液50μにつき同様に操作すると
尿素の場合と同様の結果を得る。実施例 ２
メチル尿素液50μにつき実施例１と同様に操
作し、波長520nmで吸光度を測定する。メチル尿
素は0.01〜0.3μＭの範囲において吸光度と直線
関係にある。 If the same procedure is performed for 50μ of phenyl urea solution, the same results as for urea will be obtained. Example 2 The same procedure as in Example 1 is carried out using 50μ of methyl urea solution, and the absorbance is measured at a wavelength of 520 nm. Methylurea has a linear relationship with absorbance in the range of 0.01 to 0.3 μM.
エチル尿素溶液あるいはシトルリン溶液につい
て同様に操作すると、メチル尿素の場合と同様の
結果を得る。 If the same procedure is performed with ethyl urea solution or citrulline solution, the same results as in the case of methyl urea are obtained.
実施例 ３
試薬３の調製と定量
０―フタルアルデヒド1.34ｇを50％エタノーに
溶かし25mlとする。Example 3 Preparation and quantitative determination of reagent 3 Dissolve 1.34 g of 0-phthalaldehyde in 50% ethanol to make 25 ml.
尿素溶液50μに試薬１を２mlおよび2N塩酸
２mlを加え、37℃で10分間加温する。これに試薬
３を100μ加え、波長520nmにおいて37℃、３
分間の増加する吸光度の値を測定する。この方法
により尿素0.05〜1.5μＭの範囲において尿素量
と吸光度の変化は直線関係にあり、予め作成した
検量線から尿素が定量される。 Add 2 ml of Reagent 1 and 2 ml of 2N hydrochloric acid to 50 μl of urea solution, and heat at 37°C for 10 minutes. Add 100μ of reagent 3 to this, and at 37℃ at a wavelength of 520nm,
Measure the value of increasing absorbance for minutes. With this method, there is a linear relationship between the amount of urea and the change in absorbance in the range of 0.05 to 1.5 μM of urea, and urea can be quantified from a calibration curve prepared in advance.
実施例 ４
シトルリン溶液50μにつき実施例３と同様に
操作し、波長520nmにおける吸光度の変化を測定
する。シトルリンは0.02〜0.4μＭの範囲におい
て吸光度の変化と直線関係にある。Example 4 A citrulline solution of 50μ is operated in the same manner as in Example 3, and the change in absorbance at a wavelength of 520 nm is measured. Citrulline has a linear relationship with the change in absorbance in the range of 0.02 to 0.4 μM.
メチル尿素あるいはエチル尿素溶液を用い、同
様に操作するとシトルリンの場合と同様の結果を
得る。 If a methylurea or ethylurea solution is used and the same procedure is performed, the same results as for citrulline will be obtained.
Claims (1)
ルエチレンジアミンおよび０―フタルアルデヒド
を作用させて反応液の吸光度を測定することを特
徴とする尿素およびその誘導体の定量法。1. A method for quantifying urea and its derivatives, which comprises reacting a sample with N-(1-naphthyl)-N'-diethylethylenediamine and 0-phthalaldehyde and measuring the absorbance of the reaction solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14281578A JPS5569038A (en) | 1978-11-21 | 1978-11-21 | Determining method for urea and its derivative |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14281578A JPS5569038A (en) | 1978-11-21 | 1978-11-21 | Determining method for urea and its derivative |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5569038A JPS5569038A (en) | 1980-05-24 |
| JPS6142826B2 true JPS6142826B2 (en) | 1986-09-24 |
Family
ID=15324262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14281578A Granted JPS5569038A (en) | 1978-11-21 | 1978-11-21 | Determining method for urea and its derivative |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5569038A (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59142930A (en) * | 1983-01-28 | 1984-08-16 | 田中精機株式会社 | Sheet pasting mechanism of address sheet pasting machine |
| EP2463380A1 (en) * | 2010-12-07 | 2012-06-13 | Cytonet GmbH & Co. KG | Method for isolating urea by removing undesired CO2 |
| KR101867375B1 (en) | 2010-08-20 | 2018-06-14 | 프로메테라 바이오사이언시즈 에스.에이./엔.브이. | Method for isolating urea while removing objectionable co2 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5119590A (en) * | 1974-07-05 | 1976-02-16 | Etsuchi Jungu Deibitsudo | Ryudotaino nyosoganryono hishokubunsekiho |
-
1978
- 1978-11-21 JP JP14281578A patent/JPS5569038A/en active Granted
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5119590A (en) * | 1974-07-05 | 1976-02-16 | Etsuchi Jungu Deibitsudo | Ryudotaino nyosoganryono hishokubunsekiho |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5569038A (en) | 1980-05-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Peterson | Improved spectrophotometric procedure for determination of serum iron using 4, 7-diphenyl-1, 10-phenanthroline | |
| Eggleton et al. | The estimation of creatine and of diacetyl | |
| Koepsell et al. | Microdetermination of pyruvic and α-ketoglutaric acids | |
| Gilman et al. | A qualitative color test for the Grignard reagent | |
| CN103698324B (en) | Air formaldehyde self testing kit | |
| Long | A general method for the estimation of α-keto-acids, and its application to α-keto-acid metabolism in pigeon brain | |
| JPS6142826B2 (en) | ||
| JP3312793B2 (en) | Arylazochromoionophores and methods for measuring calcium ions using the same | |
| WO2013189265A1 (en) | Method, system, and kit for detecting potassium ion concentration | |
| Neidig et al. | Simultaneous estimation of threonine and serine | |
| Cleary et al. | A simplified procedure for the measurement of total hydroxyproline in urine | |
| CN104459008A (en) | Rapid detection kit for protein and using method of kit | |
| Mathers et al. | Spectrophotometric determination of methoxyl | |
| Grogan et al. | Microdetermination of chromium in small samples of various biological media | |
| The et al. | Determination of phenylpyruvic acid in the urine of patients with oligophrenia phenylpyruvica | |
| SU567136A1 (en) | Method of evaluation of bile acid content of bile | |
| WO1994023061A1 (en) | Method of determining sodium ion | |
| CN101762543A (en) | Reagent (kit) for diagnosing/determining alcohol and method for determining concentration of alcohol | |
| Awad | Microdetermination of α-amino acids by spectrophotometric and titrimetric methods | |
| Borcherdt et al. | Tautomerization of an optically active azomethine | |
| SU1116391A1 (en) | Method of photometric determination of vanadium | |
| SU725022A1 (en) | Method of quantitative determining of nitroglycerin in medicinal preparations | |
| Kaselis et al. | Modified colorimetric assay of pteroylglutamic acid | |
| SU138087A1 (en) | Method for quantitative determination of white streptocide | |
| JPS5844210B2 (en) | Highly sensitive and simple colorimetric determination method for acetoacetic acid in blood |