JPH09154598A - Test piece for measuring activity of lipase and its production - Google Patents
Test piece for measuring activity of lipase and its productionInfo
- Publication number
- JPH09154598A JPH09154598A JP7346597A JP34659795A JPH09154598A JP H09154598 A JPH09154598 A JP H09154598A JP 7346597 A JP7346597 A JP 7346597A JP 34659795 A JP34659795 A JP 34659795A JP H09154598 A JPH09154598 A JP H09154598A
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- Japan
- Prior art keywords
- lipase
- test piece
- substrate
- layer
- measuring
- 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.)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】 液体試料、特に血清、血漿中の
リパーゼ活性を測定するための試験片とその作製方法に
関する。TECHNICAL FIELD The present invention relates to a test piece for measuring lipase activity in a liquid sample, particularly serum or plasma, and a method for producing the same.
【０００２】リパーゼ（トリアシルグリセリンアシルヒ
ドラーゼ；ＥＣ３．１．１．３）は、油滴と水性相間の
界面に乳化状態で存在する長鎖脂肪酸のトリグリセリド
を加水分解する酵素であり、急性膵炎・膵頭部癌などの
特定の疾患で血清中のリパーゼ活性が上昇する。早期診
断的価値は、アミラーゼ測定に劣るが、発病７〜１０日
以内でアミラーゼ活性がすでに正常に復帰した後でもリ
パーゼ活性は上昇を示すので経過判定に役立つ。アミラ
ーゼと合わせて測定することにより、さらに的確な膵疾
患診断が可能になる。[0002] Lipase (triacylglycerin acylhydrase; EC 3.1.1.3) is an enzyme that hydrolyzes triglycerides of long-chain fatty acids existing in an emulsified state at the interface between oil droplets and an aqueous phase, and acute pancreatitis. -In certain diseases such as pancreatic head cancer, serum lipase activity is increased. Although its early diagnostic value is inferior to that of amylase measurement, lipase activity shows an increase even after amylase activity has already returned to normal within 7 to 10 days after onset of illness, which is useful for judgment of progress. By measuring it together with amylase, more accurate diagnosis of pancreatic disease becomes possible.
【０００３】 これまでに公知のリパーゼ活性測定法は
種々あるが、代表的な測定方法を以下に記載する。 （１）比濁法 トリグリセライド、おもにオリーブ油、または高級脂肪
酸のグリセロールエステルの懸濁液を基質として酵素反
応をさせ、その濁りの減少を吸光度で測定する方法であ
る。しかし、この方法は、濁りの減少とリパーゼ活性が
必ずしも一致せず乳化検体では吸光度が逆に上昇する場
合がある。また、リパーゼ活性が低い場合は感度、再現
性ともに悪いなどの欠点がある。 （２）発色法１ フェノール、ｐ−ニトロフェノール、α−ナフトールな
どのフェノール類の長鎖脂肪酸エステルを基質とし、リ
パーゼの作用により生じるフェノール類を発色させ比色
定量する方法である。［Ａ．Ｓａｉｆｅｒ．ｅｔ ａ
ｌ． Ｃｌｉｎ．Ｃｈｅｍ．７ １７８〜１８４（１９
６１）、Ｊ．Ｆ．Ｗｈｉｔａｋｅｒ． Ｃｌｉｎｉｃａ
Ｃｈｉｍｉｃａ ａｃｔａ ４４ １３３（１９７
３）］ しかし、この方法も（１）の方法と比較すると同一試薬
で測定する際の同時再現性の向上が見られたものの、試
薬調製が煩雑であるため試薬調整を毎回同様に行うこと
は困難であるなどの欠点がある。 （３）発色法２ 特公平６−８７８００号に記載されているリパーゼ視覚
的定量用基質を用いた方法は、リパーゼによりリパーゼ
基質が分解されると発色基が遊離されるので、その発色
強度がリパーゼ活性に直接反映されるものであり、優れ
た方法である。ただし、このリパーゼ基質を用いた溶液
系のリパーゼ活性測定法においては、反応液中のリパー
ゼ基質の色が濃いオレンジ色であること、リパーゼ基質
の水不溶性の性質で濁りが生じバックグランドが高いこ
となどから有効な測定レンジが得られていない。また、
リパーゼ基質が水不溶性であることから、基質乳化溶液
を均一に、しかも毎回同様に作製することは困難であ
り、リパーゼ活性測定法の正確さが問題である。There are various known methods for measuring lipase activity, and typical measuring methods are described below. (1) Nephelometry This is a method in which a suspension of triglyceride, mainly olive oil, or a glycerol ester of a higher fatty acid is used as a substrate for an enzymatic reaction, and the decrease in turbidity is measured by absorbance. However, in this method, the decrease in turbidity and the lipase activity do not always coincide with each other, and the absorbance of the emulsified sample may increase. Further, when the lipase activity is low, there are drawbacks such as poor sensitivity and reproducibility. (2) Coloring Method 1 This is a method for colorimetrically determining the color of phenols produced by the action of lipase using a long-chain fatty acid ester of phenols such as phenol, p-nitrophenol, α-naphthol as a substrate. [A. Saifer. et a
l. Clin. Chem. 7 178-184 (19
61), J. F. Whitaker. Clinica
Chimica acta 44 133 (197)
3)] However, although this method also showed an improvement in simultaneous reproducibility when measuring with the same reagent as compared with the method of (1), the reagent preparation is complicated, so that it is not possible to perform reagent adjustment in the same manner every time. There are drawbacks such as difficulty. (3) Coloring Method 2 In the method using a substrate for lipase visual quantification described in Japanese Examined Patent Publication No. 6-87800, since the chromogenic group is released when the lipase substrate is decomposed by lipase, its coloring intensity is This is an excellent method because it is directly reflected in the lipase activity. However, in the solution-based method for measuring lipase activity using this lipase substrate, the color of the lipase substrate in the reaction solution should be dark orange, and the water-insoluble nature of the lipase substrate should cause turbidity and a high background. A valid measurement range has not been obtained. Also,
Since the lipase substrate is insoluble in water, it is difficult to prepare a substrate emulsion solution uniformly and in the same manner each time, and the accuracy of the lipase activity measurement method is a problem.
【０００４】そこで、迅速性及び簡易性のために、リパ
ーゼ測定用試薬を濾紙のような担体に含浸し、試験片と
して作製したところ、濁りによる影響が解消され、基質
溶液の調製の必要性もなくなった。しかも、測定レンジ
の拡大及び正確性の向上も見られた。Therefore, for the purpose of quickness and simplicity, a carrier such as filter paper was impregnated with a reagent for measuring lipase to prepare a test piece, and the influence of turbidity was eliminated, and the necessity of preparing a substrate solution was also required. lost. Moreover, the measurement range was expanded and the accuracy was improved.
【０００５】しかし、試験片にすることにより、経時的
なバックグランドの着色という問題が起こった。この理
由は、以下のようである。However, the use of the test piece causes a problem of background coloration over time. The reason for this is as follows.
【０００６】通常のリパーゼ活性の測定は、リパーゼの
至適ｐＨが弱アルカリ性からアルカリ性にあることから
ｐＨ７．５〜ｐＨ９．０で行われる。この条件で、リパ
ーゼ基質をｎ−プロパノールに溶解し、その他の試薬
（胆汁酸、コリパーゼ、緩衝物質など）を混合し１層の
試験片を作製してきた。しかし、ｐＨ７．５〜ｐＨ９．
０においては、リパーゼ基質液を含む溶液作製中および
試験片の保存中に、経時的にリパーゼ基質の分解が促進
された。Ordinary lipase activity is measured at pH 7.5 to pH 9.0 because the optimum pH of lipase is weakly alkaline to alkaline. Under this condition, a lipase substrate was dissolved in n-propanol and other reagents (bile acid, colipase, buffer substance, etc.) were mixed to prepare a one-layer test piece. However, pH 7.5-pH 9.
In No. 0, decomposition of the lipase substrate was promoted with time during preparation of the solution containing the lipase substrate solution and during storage of the test piece.
【０００７】また、試験片を作製するためには、リパー
ゼ基質を均一に溶解した試薬含浸液を作製する必要があ
る。リパーゼ基質は、脂質であり水溶性の溶媒には溶解
しない。そこで、溶媒として、通常ｎ−プロパノールの
ような有機溶媒を用いる。ｎ−プロパノールなどのアル
コール性溶媒は、水溶性溶媒との混合が任意に可能であ
り、また安価で汎用性に優れた溶媒である。しかし、リ
パーゼ基質のエステル結合部分と経時的にエステル交換
反応又はそれに類似した反応を起こして、経時的にリパ
ーゼ基質の分解を促進するという欠点があった。Further, in order to prepare a test piece, it is necessary to prepare a reagent impregnating solution in which a lipase substrate is uniformly dissolved. The lipase substrate is a lipid and is insoluble in a water-soluble solvent. Therefore, an organic solvent such as n-propanol is usually used as the solvent. An alcoholic solvent such as n-propanol can be arbitrarily mixed with a water-soluble solvent, is inexpensive and has excellent versatility. However, it has a drawback that it causes an ester exchange reaction or a similar reaction with the ester bond portion of the lipase substrate over time to accelerate the decomposition of the lipase substrate over time.
【０００８】試験片を作製する際の乾燥行程で、大部分
のｎ−プロパノールなどの有機溶媒は揮発するが、完全
に除くことは困難である。通常、試験片内に僅かに残
る。試験片内の僅かなｎ−プロパノールなどの有機溶媒
は、試薬片保存中にリパーゼ基質と接することになるの
で、リパーゼ基質と反応する物質であることは望ましく
ない。そこで使用する有機溶媒は、リパーゼ基質との反
応性ができるだけ少ないものが望まれる。Most of the organic solvent such as n-propanol is volatilized during the drying process in producing the test piece, but it is difficult to completely remove it. It usually remains slightly in the test piece. Since a small amount of an organic solvent such as n-propanol in the test piece comes into contact with the lipase substrate during storage of the reagent piece, it is not desirable that the substance reacts with the lipase substrate. The organic solvent used therefor is desired to have as little reactivity as possible with the lipase substrate.
【０００９】[0009]
【発明が解決しようとする課題】 本発明の目的は、上
記に示すように液体試料中のリパーゼ活性を測定する際
に、リパーゼ測定用試験片の経時的な着色を防ぎ、また
リパーゼ測定用試験片の作製中のリパーゼ基質の分解を
防ぐことによって、正確にリパーゼ活性を測定し、臨床
検査の分野で有用なリパーゼの活性測定法を提供するこ
とにある。DISCLOSURE OF THE INVENTION An object of the present invention is to prevent coloration of a lipase-measuring test piece over time when measuring the lipase activity in a liquid sample as described above, and to carry out a lipase-measuring test. It is an object of the present invention to provide a method for measuring lipase activity that is useful in the field of clinical examination, by accurately measuring lipase activity by preventing decomposition of a lipase substrate during preparation of a piece.
【００１０】[0010]
【課題を解決するための手段】 本発明は、液体試料中
のリパーゼ活性を測定するための試験片において、リパ
ーゼ基質を含有する層とリパーゼ基質を含有しない層の
２層からなることを特徴とするリパーゼ測定用試験片で
ある。Means for Solving the Problems The present invention is a test piece for measuring lipase activity in a liquid sample, which comprises two layers, a layer containing a lipase substrate and a layer not containing a lipase substrate. It is a test piece for measuring lipase.
【００１１】さらに、リパーゼ基質を含有する層におけ
るｐＨが、３．０〜５．０であり、リパーゼ基質を含有
しない層におけるｐＨが、７．５〜９．０であることを
特徴とし、２層それぞれに緩衝物質を含有することを特
徴とする。そして、該緩衝物質の最終的なモル濃度比
が、 （リパーゼ基質を含有する層）：（リパーゼ基質を含有
しない層）＝１：５〜１：１５ であることを特徴とし、液体試料点着後の反応時ｐＨ
は、ｐＨ７．５〜９．０になるリパーゼ測定用試験片で
ある。Further, the layer containing the lipase substrate has a pH of 3.0 to 5.0, and the layer not containing the lipase substrate has a pH of 7.5 to 9.0. It is characterized in that each layer contains a buffer substance. The final molar concentration ratio of the buffer substance is (layer containing lipase substrate) :( layer not containing lipase substrate) = 1: 5 to 1:15. PH during subsequent reaction
Is a lipase measurement test piece having a pH of 7.5 to 9.0.
【００１２】また、リパーゼ基質を含有する酸性層を作
製するための試薬液において、エーテル系溶媒を用いる
ことを特徴とするリパーゼ測定用試験片の作製方法であ
る。[0012] A method for producing a lipase-measuring test piece is characterized in that an ether solvent is used in a reagent solution for producing an acidic layer containing a lipase substrate.
【００１３】[0013]
【発明の実施の形態】 本発明においては、液体試料を
試験片上にスポットすることにより油状リパーゼ基質の
エマルジョンがその場で生成され、油液界面でリパーゼ
の活性が発揮される。BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, an emulsion of an oily lipase substrate is generated in situ by spotting a liquid sample on a test piece, and lipase activity is exhibited at the oil-liquid interface.
【００１４】リパーゼ基質を含有する層におけるｐＨ
は、３．０〜５．０であり、リパーゼ基質を含有しない
層におけるｐＨは、７．５〜９．０であることが好まし
い。そして、液体試料点着後の反応時ｐＨは、ｐＨ７．
５〜９．０になることが好ましい。それは、２層それぞ
れの緩衝物質の濃度に差を付けることで、反応時試薬が
混ざるとリパーゼの至適ｐＨであるｐＨ７．５〜ｐＨ
９．０にすることができる。リパーゼ基質を含有する酸
性層及びリパーゼ基質を含有しないアルカリ層が共に緩
衝物質を含有し、該緩衝物質の最終的なモル濃度比が、
１：５〜１：１５であることが好ましい。緩衝物質は、
何でもよい。PH in layers containing lipase substrate
Is 3.0 to 5.0, and the pH of the layer containing no lipase substrate is preferably 7.5 to 9.0. The reaction pH after the liquid sample is spotted is pH 7.
It is preferably 5 to 9.0. The difference is that the concentration of the buffer substance in each of the two layers is different, so that when the reagents are mixed during the reaction, the optimum pH of the lipase is pH 7.5-pH.
It can be 9.0. The acidic layer containing the lipase substrate and the alkaline layer not containing the lipase substrate both contain a buffer substance, and the final molar concentration ratio of the buffer substance is
It is preferably 1: 5 to 1:15. The buffer substance is
Anything is fine.
【００１５】リパーゼ基質としては、すでに公知であ
り、種々のものが用いられるが、長鎖脂肪酸のモノグリ
セライド、長鎖脂肪酸の１，２−ジグリセライド、長鎖
脂肪酸のトリグリセライド、ポリエチレングリセロール
長鎖脂肪酸エステル（１，２−ジオレオイルグリセロー
ルなど）、１，２−ジグリセリド−Ｄ−ニトロフェノー
ルラウリン酸エステル、三酪酸ジメチルカプロールの脂
肪酸エステル、α−ナフチルパルミテートの脂肪酸エス
テル、１，２−ｏ−ジラウリル−レック−グリセロ−３
−グルタル酸−モノエステルなどがあり、特に比色定量
用基質を用いた場合効果が大きい。また、より長鎖の脂
肪酸を用いることで、リパーゼの特異性が向上するので
好ましい。ただし、本発明は、上記リパーゼ基質に限定
されるものではない。As the lipase substrate, various substances are already known and various ones are used, and long-chain fatty acid monoglyceride, long-chain fatty acid 1,2-diglyceride, long-chain fatty acid triglyceride, polyethyleneglycerol long-chain fatty acid ester ( 1,2-dioleoylglycerol, etc.), 1,2-diglyceride-D-nitrophenol lauric acid ester, dimethylcaprol tributyric acid fatty acid ester, α-naphthyl palmitate fatty acid ester, 1,2-o-dilauryl -REC-Glycero-3
-Glutaric acid-monoester, etc., and it is particularly effective when a colorimetric substrate is used. Further, it is preferable to use a longer-chain fatty acid because the lipase specificity is improved. However, the present invention is not limited to the above lipase substrate.
【００１６】また、胆汁酸、コリパーゼ（リパーゼ活性
化蛋白）、緩衝物質を任意の層に含むことができる。Further, bile acid, colipase (lipase activating protein) and buffer substance may be contained in any layer.
【００１７】エーテル系溶媒は、例えば、１，４−ジオ
キサン、１，３−ジオキサン、テトラヒドロフラン、ジ
メチルエーテルなどが挙げられるが、特に１，４−ジオ
キサンが好ましい。Examples of the ether solvent include 1,4-dioxane, 1,3-dioxane, tetrahydrofuran and dimethyl ether, and 1,4-dioxane is particularly preferable.
【００１８】[0018]
【実施例】以下に、実施例を示すが、本発明はこれに限
定されるものではない。 本発明によるリパーゼ測定用乾式試験片の作製 （Ａ液：リパーゼ基質を含む層） リパーゼ基質： １．５ ｍｍｏｌ／ｍ２ １，２−ｏ−ジラウリル−レック−グリセロ−３ −グルタル酸−（６’−メチルレゾルフィン） −エステル （ベーリンガー・マンハイム製） １，４−ジオキサン（和光純薬製） ４５．０ ｇ／ｍ２ 酒石酸緩衝剤（ｐＨ４．０） １．５ ｍｍｏｌ／ｍ２ タウロデオキシコール酸ナトリウム ７．５ ｍｍｏｌ／ｍ２ （アルドリッチ製） コリドン Ｋ９０（ＢＡＳＦ製） １５．０ ｇ／ｍ２ （Ｂ液：リパーゼ基質を含まない層） トリス緩衝剤（ｐＨ８．５） １５．０ ｍｍｏｌ／ｍ２ コリパーゼ（フナコシ製） ９．８ ｍｇ／ｍ２ タウロデオキシコール酸ナトリウム ７．５ ｍｍｏｌ／ｍ２ （アルドリッチ製） コリドン Ｋ９０（ＢＡＳＦ製） １５．０ ｇ／ｍ２ 上記の処方に従いＡ液を試薬保持層に含浸させ乾燥させ
る。また、Ｂ液を試薬層に塗布し乾燥させた。上記試薬
保持層と試薬層を０．１％ＴｒｉｔｏｎＸ−１００水溶
液を用いラミネートした後再び乾燥し試験片を作製し
た。EXAMPLES Examples will be shown below, but the present invention is not limited thereto. Preparation of lipase dry test strip for measuring according to the present invention: lipase substrate (A liquid layer containing the lipase substrate): 1.5 mmol / m 2 1,2 -o- dilauryl - REC - glycero - 3 - glutarate - (6 '-Methylresorufin) -ester (Boehringer Mannheim) 1,4-dioxane (Wako Pure Chemical Industries) 45.0 g / m 2 Tartrate buffer (pH 4.0) 1.5 mmol / m 2 Taurodeoxycol Sodium acid salt 7.5 mmol / m 2 (manufactured by Aldrich) Kollidon K90 (manufactured by BASF) 15.0 g / m 2 (solution B: layer not containing lipase substrate) Tris buffer (pH 8.5) 15.0 mmol / m 2 colipase (manufactured by Funakoshi) 9.8 mg / m 2 sodium taurodeoxycholate 7.5 mmol / m 2 (manufactured by Aldrich) Kollidon K 90 (manufactured by BASF) 15.0 g / m 2 According to the above formulation, the solution A is impregnated into the reagent holding layer and dried. Further, the solution B was applied to the reagent layer and dried. The reagent holding layer and the reagent layer were laminated with a 0.1% Triton X-100 aqueous solution and then dried again to prepare a test piece.
【００１９】[0019]
【比較例】 従来のリパーゼ測定用乾式試験片の作製 リパーゼ基質： １．５ ｍｍｏｌ／ｍ２ １，２−ｏ−ジラウリル−レック−グリセロ−３ −グルタル酸−（６’−メチルレゾルフィン） −エステル （ベーリンガー・マンハイム製） ｎ−プロパノール（和光純薬製） ４５．０ ｇ／ｍ２ トリス緩衝剤（ｐＨ７．５） ７．５ ｍｍｏｌ／ｍ２ タウロデオキシコール酸ナトリウム １５．０ ｍｍｏｌ／ｍ２ （アルドリッチ製） コリパーゼ（フナコシ製） ９．８ ｍｇ／ｍ２ コリドン Ｋ９０（ＢＡＳＦ製） １５．０ ｇ／ｍ２ 上記の処方で、実施例と同様に試験片を作製した。Preparation lipase substrate of the comparative example a conventional lipase dry test strip for measuring: 1.5 mmol / m 2 1,2- o- dilauryl - REC - glycero - 3 - glutarate - (6'-methyl resorufin) - Ester (manufactured by Boehringer Mannheim) n-propanol (manufactured by Wako Pure Chemical Industries, Ltd.) 45.0 g / m 2 Tris buffer (pH 7.5) 7.5 mmol / m 2 sodium taurodeoxycholate 15.0 mmol / m 2 (Manufactured by Aldrich) Colipase (manufactured by Funakoshi) 9.8 mg / m 2 Kollidon K90 (manufactured by BASF) 15.0 g / m 2 A test piece was prepared in the same manner as in the example with the above formulation.
【００２０】実施例、比較例で作製した試験片における
経時的なバックグランドの着色を測定した。試料に蒸留
水を用い、蒸留水点着後１分後の５７５ｎｍにおける反
射率を分光反射率計で測定し、下記のクベルカームンク
の式より、Ｋ／Ｓ値に換算した。 クベルカームンクの式 Ｋ／Ｓ＝（１−Ｒ）２／２Ｒ Ｋ：定数 Ｓ：散乱係数 Ｒ：反射率（％）／１００The background coloration of the test pieces prepared in Examples and Comparative Examples was measured. Distilled water was used as a sample, and the reflectance at 575 nm one minute after spotting distilled water was measured with a spectroscopic reflectometer and converted into a K / S value by the following Kubelker-Munk equation. Kubelker-Munk's equation K / S = (1-R) 2 / 2R K: constant S: scattering coefficient R: reflectance (%) / 100
【００２１】４０℃保存のＫ／Ｓ値の変化を示す結果
を、表１に示した。 （ｎ＝３の平均値）The results showing the change in K / S value at 40 ° C. storage are shown in Table 1. (Average value of n = 3)
【００２２】[0022]
【表１】 [Table 1]
【００２３】本発明の試験片は、従来試験片と比較し経
時的なバックグランド着色が明らかに減少していること
がわかる。It can be seen that the background coloration of the test piece of the present invention is clearly reduced as compared with the conventional test piece.
【００２４】[0024]
【発明の効果】本発明のリパーゼ測定用試験片は、試薬
層を２層に分け、それぞれのｐＨを３．５〜５．０およ
び７．５〜９．０にし、リパーゼ基質の溶媒をｎ−プロ
パノールからエーテル系溶媒に変更することにより、試
験片作製中のリパーゼ基質の分解を軽減するだけでな
く、試験片の保存中のリパーゼ基質の分解も軽減し、バ
ックグランド着色を防ぐことができた。INDUSTRIAL APPLICABILITY In the test piece for measuring lipase of the present invention, the reagent layer is divided into two layers, the pH of each is set to 3.5 to 5.0 and 7.5 to 9.0, and the solvent for the lipase substrate is n. -By changing from propanol to ether solvent, not only can the degradation of the lipase substrate during test piece preparation be reduced, but also the degradation of the lipase substrate during storage of the test piece can be reduced, and background coloring can be prevented. It was
Claims (6)
めの試験片において、リパーゼ基質を含有する層とリパ
ーゼ基質を含有しない層の２層からなることを特徴とす
るリパーゼ活性測定用試験片。1. A test strip for measuring lipase activity in a liquid sample, which comprises two layers, a layer containing a lipase substrate and a layer not containing a lipase substrate.
が、３．０〜５．０であることを特徴とする請求項１に
記載のリパーゼ活性測定用試験片。2. pH in a layer containing a lipase substrate
Is 3.0 to 5.0, The test piece for measuring lipase activity according to claim 1, wherein
Ｈが、７．５〜９．０であることを特徴とする請求項１
〜２、いずれかに記載のリパーゼ活性測定用試験片。3. p in a layer containing no lipase substrate
H is 7.5 to 9.0.
2. A test piece for measuring lipase activity according to any one of 2 to 3.
基質を含有しない層が共に緩衝物質を含有し、該緩衝物
質の最終的なモル濃度比が、 （リパーゼ基質を含有する層）：（リパーゼ基質を含有
しない層）＝１：５〜１：１５ であり、酵素反応時のｐＨが７．５〜９．０になること
を特徴とする請求項１〜３、いずれかに記載のリパーゼ
活性測定用試験片。4. A layer containing a lipase substrate and a layer not containing a lipase substrate both contain a buffer substance, and the final molar concentration ratio of the buffer substance is (the layer containing the lipase substrate) :( the lipase substrate). (Layer not containing) = 1: 5-1: 15, and the pH during the enzyme reaction is 7.5-9.0, The lipase activity measurement according to any one of claims 1-3. Test piece.
液の作製方法において、リパーゼ基質を溶解するため
に、エーテル系溶媒を用いることを特徴とするリパーゼ
活性測定用試験片の作製方法。5. A method for producing a reagent solution in a layer containing a lipase substrate, wherein an ether solvent is used to dissolve the lipase substrate, which is a method for producing a test strip for measuring lipase activity.
であることを特徴とする請求項５に記載のリパーゼ活性
測定用試験片の作製方法。6. The method for producing a test piece for measuring lipase activity according to claim 5, wherein the ether solvent is 1,4-dioxane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7346597A JPH09154598A (en) | 1995-12-01 | 1995-12-01 | Test piece for measuring activity of lipase and its production |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7346597A JPH09154598A (en) | 1995-12-01 | 1995-12-01 | Test piece for measuring activity of lipase and its production |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH09154598A true JPH09154598A (en) | 1997-06-17 |
Family
ID=18384512
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP7346597A Pending JPH09154598A (en) | 1995-12-01 | 1995-12-01 | Test piece for measuring activity of lipase and its production |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH09154598A (en) |
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| EP1992700A1 (en) | 2007-05-16 | 2008-11-19 | FUJIFILM Corporation | Method for producing dry analytical element for pancreatic lipase measurement |
| EP2003450A1 (en) | 2007-06-12 | 2008-12-17 | Fujifilm Corporation | Dry analytical element for lipase measurement |
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-
1995
- 1995-12-01 JP JP7346597A patent/JPH09154598A/en active Pending
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| EP1825264A4 (en) * | 2004-12-03 | 2009-05-13 | Idexx Lab Inc | Methods and devices for detecting pancreatic lipase |
| EP1825264A2 (en) * | 2004-12-03 | 2007-08-29 | Idexx Laboratories, Inc. | Methods and devices for detecting pancreatic lipase |
| EP1992700A1 (en) | 2007-05-16 | 2008-11-19 | FUJIFILM Corporation | Method for producing dry analytical element for pancreatic lipase measurement |
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| EP2105509A1 (en) | 2008-03-25 | 2009-09-30 | Fujifilm Corporation | Multilayer dry analytical element for pancreatic lipase measurment |
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| JP2013235014A (en) * | 2009-04-21 | 2013-11-21 | Univ Of Tokyo | Method and device for measuring number or quantity of microorganism |
| WO2013038736A1 (en) * | 2011-09-15 | 2013-03-21 | 東洋紡株式会社 | Multilayer test piece for assaying glycosylated hemoglobin and assay method using same |
| WO2013038735A1 (en) * | 2011-09-15 | 2013-03-21 | 東洋紡株式会社 | Multilayer test piece for assaying glycosylated hemoglobin and assay method using same |
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| WO2016204276A1 (en) * | 2015-06-19 | 2016-12-22 | 株式会社シノテスト | Substrate solution for lipase activity measurement, and method and measurement reagent for measurement of lipase activity in sample |
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