JPS63279800A - Method for improving sensitivity and accuracy of analysis - Google Patents
Method for improving sensitivity and accuracy of analysisInfo
- Publication number
- JPS63279800A JPS63279800A JP11487487A JP11487487A JPS63279800A JP S63279800 A JPS63279800 A JP S63279800A JP 11487487 A JP11487487 A JP 11487487A JP 11487487 A JP11487487 A JP 11487487A JP S63279800 A JPS63279800 A JP S63279800A
- Authority
- JP
- Japan
- Prior art keywords
- cyclodextrin
- reagent
- analysis
- nitrophenol
- clathrate compound
- 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.)
- Granted
Links
- 238000004458 analytical method Methods 0.000 title claims abstract description 18
- 230000035945 sensitivity Effects 0.000 title claims abstract description 14
- 238000000034 method Methods 0.000 title claims description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 15
- 150000001875 compounds Chemical class 0.000 claims abstract description 15
- 102000013142 Amylases Human genes 0.000 claims abstract description 12
- 108010065511 Amylases Proteins 0.000 claims abstract description 12
- 235000019418 amylase Nutrition 0.000 claims abstract description 12
- 239000004382 Amylase Substances 0.000 claims abstract description 11
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000000126 substance Substances 0.000 claims abstract description 3
- 229920001450 Alpha-Cyclodextrin Polymers 0.000 claims description 14
- 229940043377 alpha-cyclodextrin Drugs 0.000 claims description 14
- 230000000694 effects Effects 0.000 abstract description 15
- 239000000758 substrate Substances 0.000 abstract description 6
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 abstract description 4
- 102100022624 Glucoamylase Human genes 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 4
- 102100024295 Maltase-glucoamylase Human genes 0.000 abstract description 3
- 108010028144 alpha-Glucosidases Proteins 0.000 abstract description 3
- 238000002425 crystallisation Methods 0.000 abstract description 3
- 230000008025 crystallization Effects 0.000 abstract description 3
- 210000002966 serum Anatomy 0.000 abstract description 3
- 238000007711 solidification Methods 0.000 abstract description 3
- 230000008023 solidification Effects 0.000 abstract description 3
- 229940039227 diagnostic agent Drugs 0.000 abstract description 2
- 239000000032 diagnostic agent Substances 0.000 abstract description 2
- 210000002700 urine Anatomy 0.000 abstract description 2
- 230000008033 biological extinction Effects 0.000 abstract 1
- 239000007853 buffer solution Substances 0.000 abstract 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 15
- 238000002835 absorbance Methods 0.000 description 11
- 229920000858 Cyclodextrin Polymers 0.000 description 10
- 238000005259 measurement Methods 0.000 description 8
- IQUPABOKLQSFBK-UHFFFAOYSA-N 2-nitrophenol Chemical class OC1=CC=CC=C1[N+]([O-])=O IQUPABOKLQSFBK-UHFFFAOYSA-N 0.000 description 6
- 102000004139 alpha-Amylases Human genes 0.000 description 5
- 108090000637 alpha-Amylases Proteins 0.000 description 5
- 229940024171 alpha-amylase Drugs 0.000 description 5
- 239000007990 PIPES buffer Substances 0.000 description 3
- HFHDHCJBZVLPGP-RWMJIURBSA-N alpha-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO HFHDHCJBZVLPGP-RWMJIURBSA-N 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 108010056771 Glucosidases Proteins 0.000 description 2
- 102000004366 Glucosidases Human genes 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000009585 enzyme analysis Methods 0.000 description 2
- 238000005057 refrigeration Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000002464 Galactosidases Human genes 0.000 description 1
- 108010093031 Galactosidases Proteins 0.000 description 1
- 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 1
- 102000001696 Mannosidases Human genes 0.000 description 1
- 108010054377 Mannosidases Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 102000012086 alpha-L-Fucosidase Human genes 0.000 description 1
- 108010061314 alpha-L-Fucosidase Proteins 0.000 description 1
- 229940025131 amylases Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 1
- 229960005147 calcium acetate Drugs 0.000 description 1
- 235000011092 calcium acetate Nutrition 0.000 description 1
- 239000001639 calcium acetate Substances 0.000 description 1
- 238000009614 chemical analysis method Methods 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000003028 enzyme activity measurement method Methods 0.000 description 1
- 150000008268 fucosides Chemical class 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 229930005346 hydroxycinnamic acid Natural products 0.000 description 1
- 235000010359 hydroxycinnamic acids Nutrition 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 150000008146 mannosides Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 125000006501 nitrophenyl group Chemical group 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- NGSWKAQJJWESNS-ZZXKWVIFSA-N trans-4-coumaric acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C=C1 NGSWKAQJJWESNS-ZZXKWVIFSA-N 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- 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、分析感度及び／又は精度を向上させる方法に
関するものであって、化学分析、酵素分析、免疫分析と
いった技術分野で重用されるものである。Detailed Description of the Invention (Industrial Field of Application) The present invention relates to a method for improving analytical sensitivity and/or accuracy, which is heavily used in technical fields such as chemical analysis, enzyme analysis, and immunoanalysis. It is.
したがって本発明は、分析、測定用試薬、臨床診断薬に
応用することができる。Therefore, the present invention can be applied to analysis and measurement reagents, and clinical diagnostic agents.
（従来の技術）
従来、酵素活性を測定する場合、ニトロフェノール誘導
体を指示薬として使用する方法が知られているが、この
ニトロフェノール系指示薬を用いる方法では、酵素が最
大活性を示すｐＨにおいて完全に解離しないため、感度
の低下は避けられなかった。(Prior art) Conventionally, when measuring enzyme activity, a method is known in which a nitrophenol derivative is used as an indicator. Since it does not dissociate, a decrease in sensitivity was inevitable.
また、これらの指示薬は、その酵素活性を測定するｐＨ
付近ではｐＨのふれに対する吸光度の安定性が悪いとい
う欠点も有していた。These indicators also have a pH value that measures their enzymatic activity.
It also had the disadvantage of poor stability of absorbance against pH fluctuations in the vicinity.
これらの欠点を解決するために、酵素活性測定において
包接化合物であるシクロデキストリンを用いて分析感度
を上昇せしめる方法が開発され（特許公表公報昭５８−
５０１３５７号）、具体的には、α−シクロデキストリ
ンを使用することによって最大感度ｐＨを７．３から７
．０に変化せしめて最大感度を約５０％高め、その結果
、ヒト血清中の α−アミラーゼ活性を測定することが
可能になったことが開示されている。In order to solve these drawbacks, a method was developed that uses cyclodextrin, which is an inclusion compound, in enzyme activity measurement to increase analytical sensitivity (Patent Publication No. 1983-
501357), specifically, the maximum sensitivity pH was increased from 7.3 to 7 by using α-cyclodextrin.
．． It is disclosed that the maximum sensitivity was increased by about 50% by changing the α-amylase activity to 0, and as a result, it became possible to measure α-amylase activity in human serum.
（発明が解決しようとする問題点）
しかしながら、このシクロデキストリンを使用する方法
では、最大感度ｐＨを所望ｐＨにシフトして測定、分析
を行うためには、非常に大量のシクロデキストリンを使
用しなければならず、そうすると、体液中の成分ハいっ
た微量成分を測定するには大量のシクロデキストリンが
妨害作用をし、正確な測定ができなくなるという欠点は
避けられなし）。(Problems to be Solved by the Invention) However, in this method using cyclodextrin, a very large amount of cyclodextrin must be used in order to shift the maximum sensitivity pH to the desired pH and carry out measurement and analysis. However, in doing so, the unavoidable disadvantage is that a large amount of cyclodextrin interferes with the measurement of trace components in body fluids, making accurate measurements impossible.
また、そのために使用するシクロデキストリン含有試薬
は、室温に保存しておいても晶出固化現像が生じて保存
性が著しく劣り、使用に耐え得ない。ましてや、このよ
うな生物系試薬は室温においたのでは成分が変質するた
め冷蔵しなければならないが、冷蔵すると更に晶出固化
現像が促進され、実用上非常に問題がある。Further, even if the cyclodextrin-containing reagent used for this purpose is stored at room temperature, crystallization and solidification occur, resulting in extremely poor storage stability and making it unusable. Moreover, such biological reagents must be refrigerated because their components deteriorate if kept at room temperature, but refrigeration further promotes crystallization, solidification, and development, which is extremely problematic in practice.
（問題点を解決するための手段）
本発明は、この欠点を解決するためになされたものであ
って、各方面から広く検討した結果、特に生体成分の分
析にはシクロデキストリンの長所を無視できないとの結
論に達した。(Means for Solving the Problem) The present invention was made to solve this drawback, and as a result of extensive studies from various aspects, the advantages of cyclodextrin cannot be ignored, especially for analysis of biological components. The conclusion was reached.
そしてシクロデキストリン法を改良するために研究を重
ねた結果、その溶解度が重要なファクーターであること
をつきとめ、溶解度の高いシクロデキストリンに着目す
るにいたった。As a result of repeated research to improve the cyclodextrin method, they discovered that solubility was an important factor, and focused on cyclodextrins with high solubility.
そこで易溶性のシクロデキストリンとしてグルコシル−
α−シクロデキストリンを使用し、ｐ−ニトロフェノー
ルを指示薬に用いて α−アミラーゼを測定したところ
、感度と精度がすぐれた測定結果が得られ、しかも試薬
の保存性ｉも全く問題がないことを確認した。そして、
このすぐれた効果は、包接化合物であればすべてのもの
について奏されることも併せて確認し、この新知見を基
礎として本発明が完成されたのである。Therefore, as a readily soluble cyclodextrin, glucosyl-
When α-amylase was measured using α-cyclodextrin and p-nitrophenol as an indicator, measurement results with excellent sensitivity and accuracy were obtained, and there were no problems with the reagent's shelf life. confirmed. and,
It was also confirmed that this excellent effect is exhibited by all clathrate compounds, and the present invention was completed based on this new knowledge.
すなわち、本発明は、溶解度の高い易溶性包接化合物を
使用することを重大なポイントとする分析の感度、精度
を向上せしめる方法である。That is, the present invention is a method for improving the sensitivity and accuracy of analysis, in which the use of easily soluble clathrate compounds with high solubility is an important point.
本発明において易溶性包接化合物としては、溶解性にす
ぐれた溶けやすい包接化合物であればすべてのものが使
用でき、その例としては、グルコシル−α−シクロデキ
ストリン、マルトシル−α−シクロデキストリンといっ
たシクロデキストリンの糖誘導体が例示できる。As the easily soluble clathrate compound in the present invention, any easily soluble clathrate compound with excellent solubility can be used, examples of which include glucosyl-α-cyclodextrin and maltosyl-α-cyclodextrin. Examples include sugar derivatives of cyclodextrin.
本発明は、特にニトロフェノール誘導体を指示薬として
用いる化学的分析方法、酵素的分析方法、免疫分析方法
に広く使昂することができ、例えばニトロフェノールを
用いるヒドロキシ桂皮酸の分析のほか、ニトロフェニル
（オリゴ）グリコシドを基質とするアミラーゼ、グルコ
シダーゼの分析、ニトロフェニルガラクトシドを基質と
矛るガラクトシダーゼの分析、同じくマンノシド、フコ
シド、ホスフェートを基質として用いるマンノシダーゼ
、フコシダーゼ、ホスファターゼの分析に有利に利用す
ることができる。The present invention can be widely used in chemical analysis methods, enzymatic analysis methods, and immunoassay methods that use nitrophenol derivatives as indicators. For example, in addition to the analysis of hydroxycinnamic acid using nitrophenol, nitrophenyl ( It can be advantageously used for the analysis of amylases and glucosidases that use oligo)glycosides as substrates, the analysis of galactosidases that use nitrophenylgalactoside as a substrate, and the analysis of mannosidases, fucosidases, and phosphatases that use mannosides, fucosides, and phosphates as substrates. .
本発明による酵素分析の原理は、ｐ−ニトロフェニール
糖基質を例えばα−アミラーゼと接触せしめた後、α−
グルコシダーゼ、グルコアミラーゼ等を用いてｐ−ニト
ロフェノールとグルコースに分解し、このようにして生
成したｐ−ニトロフェノールの黄色を例えば４０５ｎｍ
の吸光度で読みとり、α−アミラーゼの活性を測定する
のである。本発明では、この測定系において易溶性包接
化合物を添加するのであるが、その添加時期は特に限定
はなく、測定系のいずれの時期でも所定の効果が得られ
、添加時期についてデリケートな操作は全く必要でない
ので、多忙をきわめる臨床現場では特に有利であって、
本発明の実用上の効果はきわめて高いものがある。The principle of the enzyme analysis according to the present invention is that after contacting a p-nitrophenyl sugar substrate with, for example, α-amylase,
It is decomposed into p-nitrophenol and glucose using glucosidase, glucoamylase, etc., and the yellow color of p-nitrophenol thus produced is measured at 405 nm, for example.
The absorbance is read and the activity of α-amylase is measured. In the present invention, the easily soluble clathrate compound is added to this measurement system, but the timing of its addition is not particularly limited, and the desired effect can be obtained at any time in the measurement system, and delicate operations regarding the timing of addition are not required. Since it is not necessary at all, it is especially advantageous in busy clinical settings.
The practical effects of the present invention are extremely high.
易溶性包接化合物は、従来法のように大量に使用する必
要は全くなく、例えばマルトシル−又はグルコシル−α
−シクロデキストリンの場合は０．０５〜１００ｍｍｏ
ｌ／ｍｆ１．好ましくは１〜２０ｍｍｏｌ／ｍＱ程度の
使用量で充分所期の目的が達成される。したがって、極
微量存在する生体成分も正確に且つ迅速に定量すること
ができる。Easily soluble clathrate compounds do not need to be used in large quantities as in conventional methods; for example, maltosyl- or glucosyl-α
-0.05 to 100 mmo for cyclodextrin
l/mf1. Preferably, a usage amount of about 1 to 20 mmol/mQ is sufficient to achieve the intended purpose. Therefore, even biological components present in trace amounts can be quantified accurately and quickly.
本発明方法を利用して実際に分析を行うに当っては、ｐ
−ニトロフェノール結合アミラーゼ基質、易溶性包接化
合物、α−グルコシダーゼ、グルコアミラーゼ等を緩衝
液に溶解してなるアミラーゼ分析試薬を予じめ調製して
おくと好都合である。When actually performing analysis using the method of the present invention, p
- It is convenient to prepare in advance an amylase analysis reagent consisting of a nitrophenol-bound amylase substrate, a readily soluble clathrate, α-glucosidase, glucoamylase, etc. dissolved in a buffer.
この試薬は、従来品とは全く異なり、長期間冷蔵保存し
ても晶出固化することがないのできわめてすぐれている
。This reagent is completely different from conventional products and is extremely superior because it does not crystallize and solidify even when stored under refrigeration for a long period of time.
例えば、この試薬を用いてヒトの血清や尿中のアミラー
ゼを分析するには、次の操作を行えばよい。先ず、試薬
をセルに入れて３７℃に加温し；これにサンプルを加え
て混合し；ｐ−ニトロフェノールの発色を４００ｎｍ前
後における吸光度で読みとり、常法にしたがってアミラ
ーゼ活性を測定すればよいのである。For example, to analyze amylase in human serum or urine using this reagent, the following procedure may be performed. First, put the reagent in a cell and warm it to 37°C; add the sample to it and mix it; read the color development of p-nitrophenol by absorbance at around 400 nm, and measure amylase activity according to the usual method. be.
本発明は、水に易溶な包接化合物を用いることによって
分析の感度、精度を向上せしめる点を特徴とするもので
ある。そのメカニズムの詳細は、今後の研究にまたねば
ならないが、一応、診断試薬中に含まれている包接化合
物とニトロフェノール誘導体によって包接錯体を形成さ
せることにより、その吸光度を上昇させ、さらに、ＰＨ
変化に対する吸光度を安定させるものと推定される。The present invention is characterized in that the sensitivity and accuracy of analysis are improved by using a clathrate compound that is easily soluble in water. The details of the mechanism will have to be studied in the future, but at least the absorbance is increased by forming an inclusion complex with the clathrate compound contained in the diagnostic reagent and the nitrophenol derivative. P.H.
It is assumed that this stabilizes the absorbance against changes.
以下、本発明の実施例について詳記する。Examples of the present invention will be described in detail below.
実施例１ シクロデキストリン誘導体による吸光度の増
加
１００ｍｍｏｌ／ ＱのＰＩＰＥＳ緩衝液（ｐ）ｌ ７
．０）、３４ ｐ ｍｏｌ／Ｑのパラ−ニトロフェノー
ルに、グルコシル−α−シクロデキストリン、またはマ
ルトシル−α−シクロデキストリンを０〜１０ｍｍｏｌ
／ Ｑの割合でそれぞれ添加し、４０５ｎｍの吸光度を
測定し、第１図の結果を得た。Example 1 Increase in absorbance by cyclodextrin derivatives 100 mmol/Q PIPES buffer (p)l 7
．． 0), 0 to 10 mmol of glucosyl-α-cyclodextrin or maltosyl-α-cyclodextrin to 34 pmol/Q para-nitrophenol.
/Q were added, and the absorbance at 405 nm was measured to obtain the results shown in FIG.
実施例２ シクロデキストリン誘導体による吸光７一
度の安定化
１００ｍｍｏｌ／ ＱのＰＩＰＥＳ緩衝液、３４ ｐ
ｍｏｌ／ Ｄのパラ二二トロフェノール、グルコシル−
α−シクロデキストリン、またはマルトシル−α−シク
ロデキストリンを各々１０ｍｍｏｌ／ｆｌ添加し、ｐＨ
変化と吸光度の変化について測定し、第２図の結果を得
た。Example 2 Absorption by Cyclodextrin Derivatives 7 Once Stabilized 100 mmol/Q PIPES Buffer, 34 p
mol/D of para-2 ditrophenol, glucosyl-
Add 10 mmol/fl of each of α-cyclodextrin or maltosyl-α-cyclodextrin, and adjust the pH
The changes in absorbance and absorbance were measured, and the results shown in Figure 2 were obtained.
実施例３ アミラーゼ活性の測定
１ ｍｍｏｌ／Ｑのベンジリデン−パラニトロフェニル
マルトへブタオシド、２００／ｍｆｉのグルコアミラー
ゼ、１００／＋ｎＲのα−グルコシダーゼ、２０ｍｎ＋
ｏｌ＃ｌのＮａＣＱ、２ｍｍｏ１／Ｑの酢酸カルシウム
を含む１００ｍｍｏｌ＃ｌのＰＩＰＥＳ緩衡液（ｐＨ７
，０）にグルコシル−α−シクロデキストリンまたはマ
ルトシル−α−シクロデキストリンを１０ｍｍｏｌ／ｆ
ｌ加えて、３７℃において、アミラーゼ活性を測定した
ところ、第３図の結果を得た。Example 3 Measurement of amylase activity 1 mmol/Q benzylidene-paranitrophenylmaltohebutaoside, 200/mfi glucoamylase, 100/+nR α-glucosidase, 20mn+
100 mmol #l PIPES buffer containing ol #l NaCQ, 2 mmol/Q calcium acetate (pH 7
, 0) with 10 mmol/f of glucosyl-α-cyclodextrin or maltosyl-α-cyclodextrin.
In addition, amylase activity was measured at 37°C, and the results shown in Figure 3 were obtained.
この結果からも明らかなように、易溶性包接化合物を使
用することによって、アミラーゼ活性を正確に且つ高感
度で測定できることが判る。As is clear from this result, by using the easily soluble clathrate compound, amylase activity can be measured accurately and with high sensitivity.
−（発明の効果）
以上、説明したように本発明の分析感度及び精８一
度を白土させる方法は、水に易溶なシクロデキストリン
誘導体を用いることにより、指示薬ニトロフェノール誘
導体の発色、およびＰＨのふれに対する安定性が極めて
増加するという著効を奏する。- (Effects of the Invention) As explained above, the analytical sensitivity and the method of making white clay according to the present invention improve the color development of the indicator nitrophenol derivative and the pH change by using a cyclodextrin derivative that is easily soluble in water. This has the remarkable effect of greatly increasing stability against vibration.
すなわち、検体中の微量の目的物質の高感度な検出を可
能とするものである。したがって、本発明は、特に各種
の微量成分の分析に有効であり、臨床診断の分野で特に
有効である。In other words, it enables highly sensitive detection of a trace amount of a target substance in a sample. Therefore, the present invention is particularly effective in analyzing various trace components, and is particularly effective in the field of clinical diagnosis.
第１図は実施例１におけるシクロデキストリン誘導体の
添加量とその吸光度の上昇との関係を示した図であり、
第２図は実施例２におけるシクロデキストリン誘導体添
加時のｐＨ変化に対する吸光度変化を示した図である。
また、第３図は実施例３におけるアミラーゼ活性測定時
における標準曲線である。FIG. 1 is a diagram showing the relationship between the amount of cyclodextrin derivative added and the increase in absorbance in Example 1, and FIG. 2 is a diagram showing the absorbance change with respect to pH change when adding the cyclodextrin derivative in Example 2. This is a diagram. Furthermore, FIG. 3 is a standard curve for measuring amylase activity in Example 3.
Claims (1)
特徴とする分析の感度及び／又は精度の向上方法。 ２、分析系が、変換可能な化学的あるいは物理的状態を
持つ指示薬、及び、所望の分析を行うに必要な成分を含
む臨床診断薬であること、を特徴とする特許請求の範囲
第１項に記載の方法。 ３、易溶性包接化合物がグルコシル−α−シクロデキス
トリン及び／又はマルトシル−α−シクロデキストリン
である特許請求の範囲第１項記載の方法。 ４、グルコシル−α−シクロデキストリン及び／又はマ
ルトシル−α−シクロデキストリンの濃度が１〜２０ｍ
ｍｏｌ／ｍｌである特許請求の範囲第２項記載の方法。 ５、指示薬がパラ−ニトロフェノールである特許請求の
範囲第１項記載の方法。 ６、診断試薬のｐＨが６〜８である特許請求の範囲第１
項記載の方法。 ７、分析対象がアミラーゼである特許請求の範囲第１項
記載の方法。[Scope of Claims] 1. A method for improving analytical sensitivity and/or accuracy, which comprises using a readily soluble clathrate compound in an analytical system. 2. Claim 1, characterized in that the analysis system is a clinical diagnostic reagent that includes an indicator with a convertible chemical or physical state and components necessary to perform the desired analysis. The method described in. 3. The method according to claim 1, wherein the easily soluble clathrate is glucosyl-α-cyclodextrin and/or maltosyl-α-cyclodextrin. 4. The concentration of glucosyl-α-cyclodextrin and/or maltosyl-α-cyclodextrin is 1 to 20 m
The method according to claim 2, wherein the amount is mol/ml. 5. The method according to claim 1, wherein the indicator is para-nitrophenol. 6. Claim 1, wherein the diagnostic reagent has a pH of 6 to 8.
The method described in section. 7. The method according to claim 1, wherein the target to be analyzed is amylase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62114874A JP2681635B2 (en) | 1987-05-13 | 1987-05-13 | Method to improve analysis sensitivity and accuracy |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62114874A JP2681635B2 (en) | 1987-05-13 | 1987-05-13 | Method to improve analysis sensitivity and accuracy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63279800A true JPS63279800A (en) | 1988-11-16 |
| JP2681635B2 JP2681635B2 (en) | 1997-11-26 |
Family
ID=14648845
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62114874A Expired - Fee Related JP2681635B2 (en) | 1987-05-13 | 1987-05-13 | Method to improve analysis sensitivity and accuracy |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2681635B2 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09220391A (en) * | 1995-12-13 | 1997-08-26 | Juki Corp | Yarn tension control device of sewing machine |
| CN106018303A (en) * | 2016-07-12 | 2016-10-12 | 安徽伊普诺康生物技术股份有限公司 | Kit for testing alpha-L-fucosidase |
| CN109239036A (en) * | 2018-09-26 | 2019-01-18 | 北京化工大学 | A kind of nitrophenol isomers detection array |
| CN110261323A (en) * | 2019-06-21 | 2019-09-20 | 广东一方制药有限公司 | A kind of on-line evaluation method of rhizoma cibotii processing procedure |
| CN114136939A (en) * | 2021-11-18 | 2022-03-04 | 江南大学 | Method for rapidly determining nutrient digestion characteristics of starch and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3087891B2 (en) | 1998-03-31 | 2000-09-11 | 東洋紡績株式会社 | Electrolyte measurement reagent composition |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57102198A (en) * | 1980-12-15 | 1982-06-25 | Toyobo Co Ltd | Measuring method of amylase activity |
| JPS58501357A (en) * | 1981-08-28 | 1983-08-18 | バクスター インターナショナル インコーポレーテッド | Enzyme analysis method for biological fluids |
| JPS59219270A (en) * | 1983-05-30 | 1984-12-10 | Wako Pure Chem Ind Ltd | Method and reagent for stabilization of tetrazolium salt with cyclodextrin |
| JPS61200101A (en) * | 1985-02-28 | 1986-09-04 | Sanraku Inc | Partially methylated cyclodextran and production thereof |
| JPS6248399A (en) * | 1985-08-29 | 1987-03-03 | Meiji Seika Kaisha Ltd | Reagent for measuring enzymatic activity |
-
1987
- 1987-05-13 JP JP62114874A patent/JP2681635B2/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57102198A (en) * | 1980-12-15 | 1982-06-25 | Toyobo Co Ltd | Measuring method of amylase activity |
| JPS58501357A (en) * | 1981-08-28 | 1983-08-18 | バクスター インターナショナル インコーポレーテッド | Enzyme analysis method for biological fluids |
| JPS59219270A (en) * | 1983-05-30 | 1984-12-10 | Wako Pure Chem Ind Ltd | Method and reagent for stabilization of tetrazolium salt with cyclodextrin |
| JPS61200101A (en) * | 1985-02-28 | 1986-09-04 | Sanraku Inc | Partially methylated cyclodextran and production thereof |
| JPS6248399A (en) * | 1985-08-29 | 1987-03-03 | Meiji Seika Kaisha Ltd | Reagent for measuring enzymatic activity |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09220391A (en) * | 1995-12-13 | 1997-08-26 | Juki Corp | Yarn tension control device of sewing machine |
| CN106018303A (en) * | 2016-07-12 | 2016-10-12 | 安徽伊普诺康生物技术股份有限公司 | Kit for testing alpha-L-fucosidase |
| CN109239036A (en) * | 2018-09-26 | 2019-01-18 | 北京化工大学 | A kind of nitrophenol isomers detection array |
| CN110261323A (en) * | 2019-06-21 | 2019-09-20 | 广东一方制药有限公司 | A kind of on-line evaluation method of rhizoma cibotii processing procedure |
| CN114136939A (en) * | 2021-11-18 | 2022-03-04 | 江南大学 | Method for rapidly determining nutrient digestion characteristics of starch and application thereof |
| CN114136939B (en) * | 2021-11-18 | 2022-12-13 | 江南大学 | Method for rapidly determining nutrient digestion characteristics of starch and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2681635B2 (en) | 1997-11-26 |
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