JPH034800A - Testing paper for determining cholesterol - Google Patents
Testing paper for determining cholesterolInfo
- Publication number
- JPH034800A JPH034800A JP14063689A JP14063689A JPH034800A JP H034800 A JPH034800 A JP H034800A JP 14063689 A JP14063689 A JP 14063689A JP 14063689 A JP14063689 A JP 14063689A JP H034800 A JPH034800 A JP H034800A
- Authority
- JP
- Japan
- Prior art keywords
- cholesterol
- aqueous solution
- paper
- peroxidase
- color former
- 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.)
- Pending
Links
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 title claims abstract description 38
- 235000012000 cholesterol Nutrition 0.000 title claims abstract description 19
- 108010089254 Cholesterol oxidase Proteins 0.000 claims abstract description 9
- 102000003992 Peroxidases Human genes 0.000 claims abstract description 7
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 7
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims abstract description 5
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims abstract description 5
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims abstract description 5
- 238000001514 detection method Methods 0.000 claims description 8
- 239000003086 colorant Substances 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 abstract description 9
- 238000004108 freeze drying Methods 0.000 abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 239000008280 blood Substances 0.000 abstract description 2
- 210000004369 blood Anatomy 0.000 abstract description 2
- 238000004587 chromatography analysis Methods 0.000 abstract description 2
- 238000004451 qualitative analysis Methods 0.000 abstract 1
- 238000004445 quantitative analysis Methods 0.000 abstract 1
- 238000002835 absorbance Methods 0.000 description 5
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 4
- GGCLNOIGPMGLDB-GYKMGIIDSA-N cholest-5-en-3-one Chemical compound C1C=C2CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 GGCLNOIGPMGLDB-GYKMGIIDSA-N 0.000 description 2
- NYOXRYYXRWJDKP-UHFFFAOYSA-N cholestenone Natural products C1CC2=CC(=O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 NYOXRYYXRWJDKP-UHFFFAOYSA-N 0.000 description 2
- 238000007598 dipping method Methods 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- NYLUYMWPXIIXDX-UHFFFAOYSA-N 3-(phenylazaniumyl)propane-1-sulfonate Chemical class OS(=O)(=O)CCCNC1=CC=CC=C1 NYLUYMWPXIIXDX-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 101710132772 Peroxidase 1 Proteins 0.000 description 1
- SXEHKFHPFVVDIR-UHFFFAOYSA-N [4-(4-hydrazinylphenyl)phenyl]hydrazine Chemical compound C1=CC(NN)=CC=C1C1=CC=C(NN)C=C1 SXEHKFHPFVVDIR-UHFFFAOYSA-N 0.000 description 1
- 150000001448 anilines Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は、コレステロール検出試験紙に関する。[Detailed description of the invention] [Industrial application field] The present invention relates to a cholesterol detection test strip.
更に詳しくは、健康の一つの指標であるコレステロール
の迅速かつ簡易な検出を可能とする検出試験紙に関する
。More specifically, the present invention relates to a detection test strip that enables rapid and simple detection of cholesterol, which is one indicator of health.
〔従来の技術〕および〔発明が解決しようとする課題〕
コレステロールの定量は、現在酵素を用いて行われてい
るが、この方法では生成したコレステノンの吸光度を測
定しているため目視では観察できず、また酵素反応に約
１時間程度を要している。[Prior art] and [Problem to be solved by the invention]
Cholesterol is currently quantified using enzymes, but this method measures the absorbance of the produced cholestenone, making it impossible to observe visually, and the enzymatic reaction takes about an hour. .
本発明の目的は、酵素法によるコレステロールの検出法
であって、目視で定量できる試験紙を提供することにあ
る。An object of the present invention is to provide a test strip that is a method for detecting cholesterol using an enzymatic method and allows for visual determination.
かかる本発明の目的は、口紙にコレステロールオキシダ
ーゼ（ＣＯＯ）、ペルオキシダーゼ（ＰＯＤ）およびＨ
，Ｏ，発色剤（ｄｏｎｏｒ）を含浸せしめたコレステロ
ール検出試験紙によって達成される。The object of the present invention is to contain cholesterol oxidase (COO), peroxidase (POD) and H
, O, is achieved by cholesterol detection test strips impregnated with a donor.
本発明に係るコレステロール検出法の定量原理は、次の
如くである。The quantitative principle of the cholesterol detection method according to the present invention is as follows.
ＣＯＯ
コレステロール＋０２ → コレステノン＋１１□０
□０Ｄ
ｄｏｎｏｒ ＋ Ｈ，Ｏ□ → ｏｘｉｄｉｚｅｄ
ｄｏｎｏｒ ＋Ｈ２０即ち、Ｈ，Ｏ，発色剤たるｄｏｎ
ｏｒを酸化し、ｏｘｉｄｉｚｅｄｄｏｎｏｒ （チノ色
素）として発色させ、その発色量を目視定量するもので
ある。COO Cholesterol +02 → Cholestenone +11□0
□0D donor + H,O□ → oxidized
donor +H20, that is, H, O, coloring agent don
or is oxidized to develop a color as oxidized donor (Chino dye), and the amount of color developed is determined visually.
Ｈ，Ｏ，発色剤としては、１種類で発色する３、３′−
ジアミノベンチジン・４ＨＣＱ、３．３’、５，５’−
テトラメチルベンチジン、３．３’、５．５’−テトラ
メチルペンチジン・２ＨＣＱや４−アミノアンチピリン
と酸化縮合して発色するＮ−スルホプロピルアニリン類
、Ｎ−２−ヒドロキシ−３−スルホプロピルアニリン類
、フェノール類などが用いられる。H, O, and 3,3'-, which produce color with one type, are used as color formers.
Diaminobenzidine 4HCQ, 3.3', 5,5'-
Tetramethylbenzidine, 3.3', 5.5'-tetramethylpentidine, N-sulfopropylanilines that develop color through oxidative condensation with 2HCQ and 4-aminoantipyrine, N-2-hydroxy-3-sulfopropyl Anilines, phenols, etc. are used.
コレステロールオキシダーゼ、ペルオキシダーゼおよび
８２０２発色剤の口紙への含浸は、定性用、定量用、ク
ロマトグラフ用などの各種口紙を、ｐＨ６〜８のリン酸
緩衝液１０＋１２．コレステロールオキシダーゼ約５〜
４０単位、ペルオキシダーゼ約０．１〜２０単位、Ｈ２
Ｏ２発色剤約１０〜２００μｇおよび水１０ｍ Ｑより
なり、好ましくは更に約１〜５％ポリビニルピロリドン
水溶液（水との合計量で１０ｍＱ）を添加してなる酵素
−発色剤水溶液中に約１０〜６０秒間浸漬し、引き上げ
て凍結乾燥させることにより行われる。To impregnate cholesterol oxidase, peroxidase, and 8202 coloring agent into the slip paper, various slip papers for qualitative, quantitative, and chromatographic use are impregnated with a phosphate buffer solution of pH 6 to 8, 10+12. Cholesterol oxidase approximately 5~
40 units, peroxidase approximately 0.1-20 units, H2
About 10 to 60 μg of an enzyme-color former is prepared by adding about 10 to 200 μg of O2 color former and 10 m This is done by dipping for seconds, pulling out and freeze drying.
本発明に係る検出試験紙を用いることにより、血中など
のコレステロール量が目視により迅速かつ簡易に定量す
ることができる。測定に要する時間に関していえば、ポ
リビニルピロリドンを反応促進剤として用いることによ
り、発色の程度は変わらないが、その時間を約半分程度
に更に短縮することができる。By using the detection test strip according to the present invention, the amount of cholesterol in blood or the like can be quickly and easily determined visually. Regarding the time required for measurement, by using polyvinylpyrrolidone as a reaction accelerator, the time can be further shortened to about half, although the degree of color development remains the same.
次に、実施例について本発明を説明する。 Next, the present invention will be explained with reference to examples.
実施例
リン酸緩衝液（ｐＨ７） １０ｍ Ｑ
コレステロールオキシダーゼ １０単位ペルオキシ
ダーゼ １単位４−アミノアンチピリン
３０μｇ２％ポリビニルピロリドン水溶液
２ｍ１２蒸留水 ８ｍ
１２上記組成の酵素−発色剤水溶液中に、０．５Ｘ５ｃ
ｍのクロマトグラフ用口紙の先端１ｃｍの部分迄を３０
秒間浸漬し、引き上げてから凍結乾燥させて試験紙を作
製した。Example phosphate buffer (pH 7) 10m Q
Cholesterol oxidase 10 units Peroxidase 1 unit 4-aminoantipyrine 30 μg 2% polyvinylpyrrolidone aqueous solution 2 m 12 Distilled water 8 m
12 In the enzyme-coloring agent aqueous solution with the above composition, add 0.5X5c
30 mm up to the 1 cm tip of the chromatography paper
A test paper was prepared by dipping it for a second, pulling it out, and freeze-drying it.
この試験紙を、それぞれ濃度４０．８０．１００．２０
０および４００μｇ／ｄｌｌのコレステロール水溶液中
に浸漬したところ、コレステロール濃度が高くなるに従
い口紙の色が増した。This test paper has a density of 40.80.100.20, respectively.
When immersed in cholesterol aqueous solutions of 0 and 400 μg/dll, the color of the mouth paper increased as the cholesterol concentration increased.
また、このようなコレステロール水溶液各３ｍＱに上記
酵素−発色剤水溶液５０μＱを添加し、波長５５４ｎｍ
における吸光度を測定したところ、第１図のグラフに・
で示されるような結果が得られ、この結果からこのよう
な濃度範囲においてはコレステロール濃度と吸光度との
間に直線関係があり、吸光度を測定することによりコレ
ステロール濃度の測定が可能であることが分かる。Further, 50 μQ of the above enzyme-coloring agent aqueous solution was added to each 3 mQ of such cholesterol aqueous solution, and a wavelength of 554 nm was added.
When we measured the absorbance at
The results shown are obtained, and this result shows that there is a linear relationship between cholesterol concentration and absorbance in this concentration range, and that it is possible to measure cholesterol concentration by measuring absorbance. .
また、各コレステロール濃度における反応時間（発色に
要する時間）を測定すると、○で示されるように約６０
秒前後であるが、ポリビニルピロリドン水溶液を添加し
ない場合には、×で示されるように約１２０秒前後であ
った。In addition, when measuring the reaction time (time required for color development) at each cholesterol concentration, approximately 60
However, when the polyvinylpyrrolidone aqueous solution was not added, the time was about 120 seconds, as shown by x.
第１図は、コレステロール濃度と吸光度および反応時間
との関係を示すグラフである。FIG. 1 is a graph showing the relationship between cholesterol concentration, absorbance, and reaction time.
Claims (1)
ダーゼおよびＨ＿２Ｏ＿２発色剤を含浸せしめてなるコ
レステロール検出試験紙。 ２、更にポリビニルピロリドンを含浸せしめた請求項１
記載のコレステロール検出試験紙。[Claims] 1. Cholesterol detection test paper made by impregnating cholesterol oxidase, peroxidase, and H_2O_2 coloring agent into paper. 2. Claim 1 further impregnated with polyvinylpyrrolidone
Cholesterol detection test strip as described.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14063689A JPH034800A (en) | 1989-06-02 | 1989-06-02 | Testing paper for determining cholesterol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14063689A JPH034800A (en) | 1989-06-02 | 1989-06-02 | Testing paper for determining cholesterol |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH034800A true JPH034800A (en) | 1991-01-10 |
Family
ID=15273291
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14063689A Pending JPH034800A (en) | 1989-06-02 | 1989-06-02 | Testing paper for determining cholesterol |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH034800A (en) |
-
1989
- 1989-06-02 JP JP14063689A patent/JPH034800A/en active Pending
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