JPH06104073B2 - Manufacturing method of test piece for body fluid component inspection - Google Patents
Manufacturing method of test piece for body fluid component inspectionInfo
- Publication number
- JPH06104073B2 JPH06104073B2 JP3280689A JP3280689A JPH06104073B2 JP H06104073 B2 JPH06104073 B2 JP H06104073B2 JP 3280689 A JP3280689 A JP 3280689A JP 3280689 A JP3280689 A JP 3280689A JP H06104073 B2 JPH06104073 B2 JP H06104073B2
- Authority
- JP
- Japan
- Prior art keywords
- test piece
- body fluid
- parts
- ink composition
- printing ink
- 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 - Lifetime
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- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】 〔発明の目的〕 (産業上の利用分野) 本発明は,試験薬,診断薬など,製剤化学および分析化
学の分野で広く利用可能な,体液成分検査用試験片の製
造方法に関する。DETAILED DESCRIPTION OF THE INVENTION Object of the Invention (Industrial field of application) The present invention provides a test piece for testing body fluid components, which can be widely used in the fields of pharmaceutical chemistry and analytical chemistry, such as test agents and diagnostic agents. It relates to a manufacturing method.
(従来の技術) 試験薬,診断薬など,製剤化学および分析化学の分野で
は,強酸,強アルカリなどの強い試薬を使用せず,比較
的おだやかな条件下で体液中の病理学的成分を測定する
ことができるため,酵素,ホルモン,抗原―抗体反応ま
たはハプテン抗体反応に関与する能力のある物質,凝固
因子などの生物学的活性たんぱく質を用いる測定法が,
特公昭45−1878号公報記載のぶどう糖検出用品などをは
じめとして広く用いられている。(Prior art) In the fields of pharmaceutical chemistry and analytical chemistry such as test drugs and diagnostic agents, pathological components in body fluids are measured under relatively mild conditions without using strong reagents such as strong acids or strong alkalis. Therefore, the assay method using biologically active protein such as enzyme, hormone, substance capable of participating in antigen-antibody reaction or hapten antibody reaction, coagulation factor, etc.
It is widely used, including the glucose detection products described in Japanese Examined Patent Publication No. 45-1878.
これらの検査用試験片は,通常,ろ紙などの吸収性材
料に検査薬組成物を含浸させ乾燥させたものを小片に裁
断し,この小片をプラスチックフィルムなどの機械的強
度の高い基材に接着剤などを用いて貼付して製造する
か,基材に,検査薬組成物を印刷して製造されてい
る。しかしながら,上記の方法は,製造工程がきわめ
て煩雑であり,コストが高くなるという欠点があった。These test pieces are usually obtained by impregnating an absorbent material such as filter paper with the test agent composition and drying it, and then cutting it into small pieces and adhering the small pieces to a substrate with high mechanical strength such as a plastic film. It is manufactured by sticking it using an agent or the like, or by printing the test drug composition on the base material. However, the above method has a drawback that the manufacturing process is extremely complicated and the cost is high.
上記の方法としては,生物学的活性たんぱく質である
酵素を用い,検査薬を水性組成物として基材に印刷する
方法(特公昭44−25953号公報など参照)や,検査薬を
水―アルコール性組成物として基材に印刷する方法(実
開昭57−79767号公報など参照)があるが,いずれも検
査薬組成物が多量の水を含むため,印刷後,常温あるい
は常温に低い低温で長時間かけて乾燥しなければなら
ず,作業性や生産性に劣る,短時間で乾燥しようとし
て,加熱乾燥すると酵素が失活してしまうという欠点が
あった。より低温短時間で乾燥させるために,酵素をn
−ブタノールなどの非水性溶媒に分散させた組成物を検
出薬として基材に印刷する方法(特開昭58−209995号公
報,特開昭60−178358号公報,特開昭61−284661号公報
など参照)も提案されているが,低温短時間とはいえ,
耐熱性の低い酵素を加熱するため,検査薬組成物中の酵
素の大部分が失活し,有効酵素量が減少するので,高価
な酵素を多量に用いなければならないという欠点があっ
た。As the above-mentioned method, an enzyme which is a biologically active protein is used and a test agent is printed on a substrate as an aqueous composition (see Japanese Patent Publication No. 44-25953), or the test agent is water-alcoholic. There is a method of printing on a substrate as a composition (see Japanese Utility Model Publication No. 57-79767, etc.). However, since the test agent composition contains a large amount of water in both cases, after printing, it can be stored at room temperature or at a low temperature lower than room temperature for a long time. It had to be dried for a long time, which was inferior in workability and productivity, and there was a drawback that the enzyme was inactivated by heating and drying in an attempt to dry in a short time. In order to dry at a lower temperature in a short time, the enzyme
A method of printing a composition dispersed in a non-aqueous solvent such as butanol on a substrate as a detection agent (JP-A-58-209995, JP-A-60-178358, JP-A-61-284661) (See, etc.) is also proposed, but even though it is low temperature and short time,
Since the enzyme having a low thermostability is heated, most of the enzyme in the test drug composition is inactivated and the amount of effective enzyme is reduced, so that there is a drawback that a large amount of expensive enzyme must be used.
(発明が解決しようとする課題) 本発明は,低温短時間で硬化乾燥できるために,検査薬
組成物中の酵素などの生物学的活性たんぱく質が失活す
ることなく,高価な生物学的活性たんぱく質を所要量使
用するだけですみ,作業性および生産性の良好な,体液
成分検査用試験片の製造方法を提供するものである。(Problems to be Solved by the Invention) Since the present invention can be cured and dried at low temperature in a short time, biologically active proteins such as enzymes in test drug compositions are not inactivated, and expensive biological activity is not caused. It is intended to provide a method for producing a test piece for inspecting body fluid components, which requires only a required amount of protein and has good workability and productivity.
(課題を解決するための手段) 本発明は,基材上に,生物学的活性たんぱく質,指示薬
および放射線硬化性化合物からなる印刷インキ組成物を
印刷し,印刷された印刷インキ組成物を活性放射線照射
により硬化することを特徴とする体液成分検査用試験片
の製造方法である。(Means for Solving the Problem) The present invention prints a printing ink composition comprising a biologically active protein, an indicator and a radiation-curable compound on a substrate, and prints the printed printing ink composition with active radiation. A method for producing a test piece for body fluid component inspection, which is characterized by being cured by irradiation.
本発明において基材としては,ろ紙,アート紙などの
紙,合成紙,不織布,織布,プラスチックフィルム,発
泡プラスチックシート,金属箔,あるいはこれらの積層
体などが用いられる。In the present invention, as the substrate, paper such as filter paper, art paper, synthetic paper, non-woven fabric, woven cloth, plastic film, foamed plastic sheet, metal foil, or a laminated body of these is used.
本発明において生物学的活性たんぱく質としては,植物
性または動物性の活性たんぱく質,あるいはこれらに架
橋剤などの化学物質をグラフトさせた形の変性体が用い
られる。このような生物学的活性たんぱく質としては,
酵素,ホルモン,抗原―抗体反応またはハプテン抗体反
応をおこす生物学的たんぱく質,凝固因子などがあり,
酵素としては,生体内とりわけ血液などの体液中,ある
いは尿中に含有される病理学的成分,例えば,カルシウ
ム,マグネシウムなどの無機質成分,グルコースなどの
グリコース,コレステロール,尿素,尿酸,クレアチニ
ン,中性脂肪,りん脂質,アンモニア,乳酸,ピルビン
酸,シアル酸,GPT,GOT,コリンエステラーゼ,ロイシン
アミノペプチターゼ,アミラーゼ,LCAT,クレアチンホス
ホキナーゼなどと反応し,指示薬の色調変化をおこすも
の,あるいは前記反応によって生成した物質がさらに他
の酵素と反応して指示薬の色調変化をおこすもの,ある
いは,これらの反応の結果,発光し,またはスペクトル
変化をおこし,可視紫外線吸収スペクトル,蛍光スペク
トルなどの分光光度計によって分析できるものが用いら
れる。また,抗原―抗体反応あるいはハプテン抗体反応
をおこす生物学的活性たんぱく質としては,特に制限は
なく,たんぱく質などの高分子物質に結合すると免疫応
答を示す物質していのハプテンや免疫グロブリンと総称
される抗体などであり,ヒト,サル,マウス,ラット,
イヌ,ウサギ,ウシ,ウマ,鳥類,爬虫類,両棲類,魚
類,円口類などから得られる免疫グロブリンを用いるこ
とができるが,これらに限定されるものではない。In the present invention, as the biologically active protein, a plant or animal active protein, or a modified form thereof in which a chemical substance such as a crosslinking agent is grafted is used. Such biologically active proteins include:
There are enzymes, hormones, antigen-antibody reactions or biological proteins that cause hapten antibody reactions, coagulation factors, etc.,
Enzymes include pathological components contained in body fluids such as blood in the body, or in urine, such as inorganic components such as calcium and magnesium, glucose such as glucose, cholesterol, urea, uric acid, creatinine, and neutral. By reacting with fats, phospholipids, ammonia, lactic acid, pyruvic acid, sialic acid, GPT, GOT, cholinesterase, leucine aminopeptidase, amylase, LCAT, creatine phosphokinase, etc., which changes the color tone of the indicator, or by the above reaction The produced substance reacts with other enzymes to change the color tone of the indicator, or as a result of these reactions, it emits light or changes in spectrum, and is detected by a spectrophotometer such as visible ultraviolet absorption spectrum or fluorescence spectrum. What can be analyzed is used. The biologically active protein that causes an antigen-antibody reaction or a hapten antibody reaction is not particularly limited, and is collectively referred to as hapten or immunoglobulin, which are substances that show an immune response when bound to a high molecular substance such as protein. Such as antibodies, humans, monkeys, mice, rats,
Immunoglobulins obtained from dogs, rabbits, cows, horses, birds, reptiles, amphibians, fishes, ornithopods can be used, but are not limited thereto.
本発明において指示薬としては,酵素還元反応により発
光したり,発色したり,色変化を示したり,スペクトル
変化をおこしたりする化合物(一成分系指示薬),ある
いは酸化還元反応により生成した化合物がさらに他の化
合物(カプラー)と結合して発光したり,発色したり,
色変化を示したり,スペクトル変化をおこしたりするよ
うな化合物(顕色剤)である。一成分系指示薬として
は,o−トルイジン,3,3′,5,5′−テトラメチルベンジ
ン,グアヤク脂,p−アニシジン,N,N−ジメチルp−フェ
ニレンジアミン,2,7−ジアミノフルオレン,2,2′−ジア
ノビス(3−エチルベンゾチアゾリン−6−スルホン
酸),2,6−ジクロロフェノール,α−ナフトール,よう
化カリウムなどがある。顕色剤としては,4−アミノアン
チピリン,3−メチル−2−ベンゾチアゾリノンヒドラゾ
ン,テトラメチルベンジジン,N,N−ジメチル−p−フェ
ニレンジアミンなどがある。カプラーとしては,1,7−ジ
ヒドロキシナフタレン,1,8−ジヒドロキシ−3,6−ジス
ルホン酸ナフタレン,8−アミノ−1,1−ナフトール−3,6
−ジスルホン酸スフタレンなどのナフトール類,ジメル
アニリン,ジエチルアニリン,N−メチル−N−ヒドロキ
シエチルアニリン,N−メチル−N−ヒドロキシエチル−
m−トルイジン,N,N−ジメチル−m−アニジンなどのア
ニリン類,フェノール,p−クロロフェノール,2,6−ジク
ロロフェノール,グアヤコール,ピロガロール,o−フェ
ニルフェノールなどのフェノール類,1−ナフトール−3,
6−ジスルホン酸,1−アミノナフタレン−5−スルホン
酸,1−アミノナフタレン−6−スルホン酸などのスルホ
ン酸,あるいはこれらのスルホン酸と高級アルキルアミ
ンとの塩類などがある。In the present invention, as the indicator, a compound (one-component indicator) that emits light, develops a color, shows a color change, or undergoes a spectrum change by an enzymatic reduction reaction, or a compound produced by a redox reaction is further included. When combined with the compound (coupler), it emits light, develops color,
It is a compound (developing agent) that exhibits a color change or a spectrum change. One-component indicators include o-toluidine, 3,3 ', 5,5'-tetramethylbenzine, guaiac butter, p-anisidine, N, N-dimethyl p-phenylenediamine, 2,7-diaminofluorene, 2 2,2'-dianobis (3-ethylbenzothiazoline-6-sulfonic acid), 2,6-dichlorophenol, α-naphthol, potassium iodide and the like. Examples of developers include 4-aminoantipyrine, 3-methyl-2-benzothiazolinone hydrazone, tetramethylbenzidine, and N, N-dimethyl-p-phenylenediamine. As couplers, 1,7-dihydroxynaphthalene, 1,8-dihydroxy-3,6-disulfonic acid naphthalene, 8-amino-1,1-naphthol-3,6
-Naphthols such as phthalene disulfonate, dimeraniline, diethylaniline, N-methyl-N-hydroxyethylaniline, N-methyl-N-hydroxyethyl-
Anilines such as m-toluidine, N, N-dimethyl-m-anidin, phenols, p-chlorophenol, 2,6-dichlorophenol, guaiacol, pyrogallol, phenols such as o-phenylphenol, 1-naphthol-3 ,
There are sulfonic acids such as 6-disulfonic acid, 1-aminonaphthalene-5-sulfonic acid and 1-aminonaphthalene-6-sulfonic acid, and salts of these sulfonic acids with higher alkylamines.
本発明において放射線硬化性化合物としては,(メタ)
アクリル酸,(メタ)アクリル酸塩類,(メタ)アクリ
ル酸アルキルエステル類,(メタ)アクリル酸グリシジ
ルエステル,(メタ)アクリル酸ヒドロキシアルキルエ
ステル類,(メタ)アクリロニトリル,(メタ)アクリ
ルアミド,スチレン,α−アルキルスチレン類,酢酸ビ
ニル,塩化ビニリデン,ビニルエーテル類,アリルアル
コールなど,あるいはこれらの置換体,ジ(メタ)アク
リレート類,トリ(メタ)アクリレート類などのポリ
(メタ)アクリレート類,ジ(メタ)アクリルアミド
類,ジアリル(イソ)フタレート,ジアリル(イソ)フ
タレートオリゴマー,ジビニルベンゼン,トリアリル化
合物など,あるいはこれらの混合物がある。In the present invention, the radiation-curable compound includes (meth)
Acrylic acid, (meth) acrylic acid salts, (meth) acrylic acid alkyl esters, (meth) acrylic acid glycidyl ester, (meth) acrylic acid hydroxyalkyl esters, (meth) acrylonitrile, (meth) acrylamide, styrene, α -Alkyl styrenes, vinyl acetate, vinylidene chloride, vinyl ethers, allyl alcohol, etc., or substitution products thereof, poly (meth) acrylates such as di (meth) acrylates, tri (meth) acrylates, di (meth) There are acrylamides, diallyl (iso) phthalate, diallyl (iso) phthalate oligomers, divinylbenzene, triallyl compounds, etc., or a mixture thereof.
本発明において,試験検査薬としての印刷インキ組成物
は,上記生物学的活性たんぱく質,指示薬,および放射
線硬化性化合物の他,本発明の体液成分検査用試験片の
性能を阻害しない範囲で,必要に応じて,吸液性および
保液性の向上のためのシリカ,カオリンなどの無機質充
填剤,pH緩衝剤,界面活性剤,酸化防止剤,吸液性およ
び保液性向上のためのカルボキシメチルセルロースナト
リウム塩,メチルセロルース,寒天,アルギン酸ナトリ
ウム,ポリアクリル酸ナトリウム,ゼラチン,でん粉な
どの水溶性ポリマー類,染顔料,重合禁止剤,光重合開
始剤,光重合促進剤,滑剤,熱可塑性樹脂,少量の水や
有機溶剤,少量の熱硬化性樹脂などの添加剤を加え,十
分混合しまたは混練することによって得られる。このよ
うにして得られた印刷インキ組成物中の指示薬の量は0.
1〜5重量%であることが好ましい。In the present invention, a printing ink composition as a test and inspection agent is required within the range that does not impair the performance of the biological fluid component test piece of the present invention, in addition to the above biologically active protein, indicator, and radiation curable compound. Depending on the type, silica, inorganic fillers such as kaolin, pH buffers, surfactants, antioxidants, carboxymethyl cellulose for improving liquid absorption and liquid retention properties for improving liquid absorption and liquid retention properties Water-soluble polymers such as sodium salt, methylcellulos, agar, sodium alginate, sodium polyacrylate, gelatin, starch, dyes and pigments, polymerization inhibitors, photopolymerization initiators, photopolymerization accelerators, lubricants, thermoplastic resins, It can be obtained by adding a small amount of water, an organic solvent, a small amount of an additive such as a thermosetting resin, and thoroughly mixing or kneading. The amount of indicator in the printing ink composition thus obtained is 0.
It is preferably from 1 to 5% by weight.
このようにして得られた試験検査薬としての印刷インキ
組成物は,単独であるいは適宜組合せて前記基材に,あ
るいは単独または組合わせたもの2種以上を前記基材の
異なった部分に,凸版印刷,平版印刷,グラビア印刷,
フレキソ印刷,スクリーン印刷などの基材の種類と印刷
インキ組成物の種類により選ばれる適切な印刷方式によ
り印刷され,基材の印刷部分に,紫外線,電子線,X−線
などの活性放射線を照射して,これを硬化させて,必要
に応じて適当な大きさに裁断して,本発明の体液成分検
査用試験片が得られる。活性放射線が紫外線である場合
には,硬化性の面から,印刷インキ組成物中に光重合開
始剤および必要に応じて光重合促進剤を含むことが好ま
しい。また,基材が活性放射線透過性を有する場合に
は,基材の印刷面と反対側から活性放射線を照射して,
印刷部分を硬化させることもできる。The printing ink composition as a test test agent thus obtained may be used alone or in appropriate combination on the substrate, or two or more of them may be used alone or in combination on different portions of the substrate. Printing, lithographic printing, gravure printing,
Printing is performed by an appropriate printing method selected according to the type of substrate and printing ink composition such as flexographic printing and screen printing, and the printed portion of the substrate is irradiated with actinic radiation such as ultraviolet rays, electron beams, and X-rays. Then, this is cured, and if necessary, cut into an appropriate size to obtain the test piece for body fluid component inspection of the present invention. When the actinic radiation is ultraviolet rays, it is preferable from the viewpoint of curability that the printing ink composition contains a photopolymerization initiator and, if necessary, a photopolymerization accelerator. When the substrate has actinic radiation transparency, the actinic radiation is applied from the side opposite to the printed surface of the substrate,
The printed portion can also be cured.
このようにして得られた体液成分検査用試験片を用い
て,生体内とりわけ血液などの体液中,あるいは尿中に
含有される病理学的成分,例えば,カルシウム,マグネ
シウム,などの無機質成分,グルコースなどのグリコー
ス,コレステロール,尿素,尿酸,クレアチニン,中性
脂肪,りん脂質,アンモニア,乳酸,ピルビン酸,シア
ル酸,GPT,GOT,コリンエステラーゼ,ロイシンアミノペ
プチターゼ,アミラーゼ,LCAT,クレアチンホスホキナー
ゼなどを検査することができる。By using the thus obtained test piece for testing body fluid components, pathological components contained in body fluid such as blood or urine in vivo, for example, inorganic components such as calcium and magnesium, glucose Glucose, cholesterol, urea, uric acid, creatinine, neutral fat, phospholipids, ammonia, lactic acid, pyruvate, sialic acid, GPT, GOT, cholinesterase, leucine aminopeptidase, amylase, LCAT, creatine phosphokinase, etc. can do.
(実施例) 以下,実施例により本発明を説明する。例中,部とは重
量部を表す。(Examples) Hereinafter, the present invention will be described with reference to Examples. In the examples, “part” means “part by weight”.
実施例1. 緩衝剤分散液Aの調整 ビニルピロリドン 125部 くえん酸一水和物 5部 くえん酸三ナトリウム二水和物 20部 2,6−ジ−tert−ブチルヒドロキシトルエン 0.07部 上記組成の混合物を,ボールミルで2日間分散させ,乳
白色の緩衝剤分散液Aを得た。Example 1. Preparation of buffer dispersion A Vinylpyrrolidone 125 parts Citric acid monohydrate 5 parts Trisodium citrate dihydrate 20 parts 2,6-Di-tert-butylhydroxytoluene 0.07 parts Mixture of the above composition Was dispersed with a ball mill for 2 days to obtain a milky white buffer dispersion liquid A.
分散液Bの調整 緩衝剤分散液A 15部 ビニルアルコール−アクリル酸塩ブロック共重合体(高
吸水性ゼル)「スミカゲルSP520」(住友化学工業
(株)製,商品名) 8部 ポリウチレングリコール6000 1部 ポリビニルピロリドン 7部 グリセリンジグリシジルエーテルジアクリレート 1部 グアヤク脂 0.8部 上記組成の混合物を,ホモジナイザーを用いて混合分散
し,分散液Bを得た。Preparation of Dispersion B Buffer Dispersion A 15 parts Vinyl alcohol-acrylate block copolymer (super absorbent gel) "Sumikagel SP520" (Sumitomo Chemical Co., Ltd., trade name) 8 parts Polyethylene glycol 6000 1 part Polyvinylpyrrolidone 7 parts Glycerin diglycidyl ether diacrylate 1 part Guayak fat 0.8 parts The mixture having the above composition was mixed and dispersed using a homogenizer to obtain a dispersion B.
酵素分散液Cの調整 りん酸緩衝液(pH7.0,1/15M) 1部 グルコースオキシダーゼ(タイプII,シグマ社製)0.2部 パーオキシダーゼ(タイプII,シグマ社製) 0.03部 ポリエチレングリコール400 1部 氷浴上で,上記りん酸緩衝液に,上記グルコースオキシ
ダーゼ,パーオキシダーゼおよびポリエチレングリコー
ルをこの順に溶解させ,やや濁りのある茶褐色の粘稠な
酵素分散液Cを得た。Preparation of Enzyme Dispersion C Phosphate buffer (pH 7.0,1 / 15M) 1 part Glucose oxidase (Type II, Sigma) 0.2 parts Peroxidase (type II, Sigma) 0.03 parts Polyethylene glycol 400 1 part The above glucose oxidase, peroxidase and polyethylene glycol were dissolved in this order in the above phosphate buffer on an ice bath to obtain a slightly turbid brownish viscous enzyme dispersion C.
印刷インキ組成物の調整 酵素分散液C2.23部を,分散液B32.8部に氷冷しながら加
え,ホモジナイザーで10分間攪拌分散させ,次いで,2−
ヒドロキシ−2−メチルプロピオフェノン0.5部を加
え,さらに5分間攪拌し,印刷インキ組成物Dを得た。Preparation of printing ink composition Add 2.23 parts of Enzyme Dispersion C to 32.8 parts of Dispersion B while ice-cooling, stir and disperse for 10 minutes with a homogenizer, then
0.5 parts of hydroxy-2-methylpropiophenone was added and stirred for 5 minutes to obtain a printing ink composition D.
体液成分検査用試験片の調整および評価 印刷インキ組成物Dを,100メッシュのポリテトラフルオ
ロエチレン製スクリーンを用いて,厚さ200μmの透明
ポリスチレンフィルムの片面に,1cm幅,印刷膜厚35μm
となるように,スクリーン印刷し,印刷物を得た。得ら
れた印刷物を,80W/cm紫外線ランプ下,6.5m/分の速度で
4回通過させることによって,フィルムの印刷側から紫
外線を照射し,印刷部分を硬化させた。この際の紫外線
の総照射量は200mJ/cm2であった。また,印刷硬化部分
にはタックはなく,印刷硬化部分のポリスチレンフィル
ム基材への密着性も良好であった。Preparation and evaluation of test pieces for body fluid component inspection Using a 100-mesh polytetrafluoroethylene screen, printing ink composition D was coated on one side of a 200 μm thick transparent polystyrene film with a 1 cm width and a printing film thickness of 35 μm.
Then, screen printing was performed to obtain a printed matter. The obtained printed matter was passed through an 80 W / cm ultraviolet lamp at a speed of 6.5 m / min four times to radiate ultraviolet rays from the print side of the film to cure the printed portion. The total irradiation dose of ultraviolet rays at this time was 200 mJ / cm 2 . In addition, there was no tack in the print-cured part, and the adhesion of the print-cured part to the polystyrene film substrate was good.
検査薬区域の大きさがたて1cm横1cmの正方形となるよう
に,得られた印刷硬化フィルムを裁断して試験片を得,
得られた試験片を,種々の濃度のグルコース水溶液に3
秒間浸漬し,取出した後,1分経過後の発色を,シアン色
側光用フィルターを用いて,反射濃度計「B−318G」
(X−Rite,Inc.製,商品名)により測定した。測定結
果を表1に示す。なお,反射濃度の数値は,それが大で
あるほど色が濃いことを示す。また,グリコース濃度0
〜500mg/dlの範囲では,反射濃度の大きな変化がみら
れ,標準色と比較して目視により明確にグルコース濃度
を測定することができる。The obtained print-cured film was cut so that the size of the test drug area was a square of 1 cm in width and 1 cm in width, and a test piece was obtained.
The obtained test pieces were applied to glucose aqueous solutions of various concentrations.
After immersing for 2 seconds and then taking it out, the color developed after 1 minute was measured using a cyan-side light filter for reflection densitometer "B-318G".
(X-Rite, Inc., trade name). The measurement results are shown in Table 1. The numerical value of the reflection density indicates that the larger the value, the darker the color. Also, the concentration of glucose is 0
In the range of up to 500 mg / dl, a large change in reflection density was observed, and the glucose concentration can be clearly measured by visual observation as compared with the standard color.
表1中,グルコース濃度の単位はmg/dlである。以下の
表でも同様である。 In Table 1, the unit of glucose concentration is mg / dl. The same applies to the table below.
また,得られた試験片の可視吸収スペクトルを測定し,
波長λ1=480nm,λ2=610nmおよびλ=740nmそれぞれに
おける吸光度Eλ1,Eλ2,およびEλ3を求め,λ2にお
ける,ベースラインに相当する吸光度を差引いた実質的
な吸光度Eλ2,sを, 式 Eλ2,s=Eλ2−(Eλ1+Eλ2)/2 に基づいて算出した。表2に算出結果を示す。Also, measure the visible absorption spectrum of the obtained test piece,
Absorbances Eλ 1 , Eλ 2 , and Eλ 3 at wavelengths λ 1 = 480 nm, λ 2 = 610 nm and λ = 740 nm, respectively, are obtained, and the actual absorbance Eλ 2 , s at λ 2 is subtracted from the absorbance corresponding to the baseline. Was calculated based on the equation Eλ 2 , s = Eλ 2 − (Eλ 1 + Eλ 2 ) / 2. Table 2 shows the calculation results.
表2から,グルコース濃度とEλ2,sとの間には直線的
な関係はないが,可視吸収スペクトルの吸光度を用いた
体液成分の濃度決定が可能であることがわかる。 From Table 2, it is understood that there is no linear relationship between the glucose concentration and Eλ 2 , s, but the concentration of the body fluid component can be determined using the absorbance of the visible absorption spectrum.
実施例2. 2,6−ジ−tert−ブチルヒドロキシトルエンの代わりに
ブチル化ヒドロキシアニソールを用い,2−ヒドロキシ−
2−メチルプロピオフェノンを用いなかった以外は,実
施例1と同様にして,印刷インキ組成物Eを得た。得ら
れた印刷インキ組成物Eを,実施例1と同様にして,透
明ポリスチレンフィルム基材に印刷し,カーテン方式電
子線照射装置(エナジーサイエンス社製)を用いて,ち
っ素雰囲気中,160kv,5mAで,線量7Mradの電子線を照射
し,印刷部分を硬化させた。この際,印刷硬化部分には
タックはなく,印刷硬化部分のポリスチレンフィルム基
材への密着性も良好であった。Example 2.Butylated hydroxyanisole was used in place of 2,6-di-tert-butylhydroxytoluene, and 2-hydroxy-
A printing ink composition E was obtained in the same manner as in Example 1 except that 2-methylpropiophenone was not used. The obtained printing ink composition E was printed on a transparent polystyrene film substrate in the same manner as in Example 1, and a curtain type electron beam irradiation device (Energy Science Co., Ltd.) was used in a fluorine atmosphere at 160 kv, The printed area was cured by irradiating it with an electron beam at a dose of 7 Mrad at 5 mA. At this time, there was no tack in the print-cured part, and the adhesion of the print-cured part to the polystyrene film substrate was good.
実施例1と同様にして,得られた印刷硬化フィルムから
試験片を得,得られた試験片を用いて,種々のグルコー
ス濃度における反射濃度を測定した。測定した結果を表
3に示す。A test piece was obtained from the obtained print-cured film in the same manner as in Example 1, and the reflection density at various glucose concentrations was measured using the obtained test piece. Table 3 shows the measured results.
なお,グルコース濃度による発色の差は,目視によって
も明瞭に区別できるものであり,グルコースの定量が可
能であった。 In addition, the difference in color development depending on the glucose concentration was clearly distinguishable by visual observation, and it was possible to quantify glucose.
実施例3. 2,6−ジ−tert−ブチルヒドロキシトルエンの代わりに
ブチル化ヒドロキシアニソールを,グアヤク脂0.8部の
代わりに4−アミノアンチピリン0.4部およびサリチル
アニリド0.4部を,それぞれ用いた以外は実施例1と同
様にして,印刷インキ組成物Fを得た。実施例1と同様
にして,透明ポリスチレンフィルムに印刷し,紫外線照
射によって硬化させ,試験片を得,シアン色側光用フィ
ルターを用い,種々のグルコース濃度における反射濃度
を測定した。測定した結果を表4に示す。Example 3. Carried out except that butylated hydroxyanisole was used instead of 2,6-di-tert-butylhydroxytoluene, and 0.4 parts of 4-aminoantipyrine and 0.4 parts of salicylanilide were used instead of 0.8 part of guaiac butter, respectively. A printing ink composition F was obtained in the same manner as in Example 1. In the same manner as in Example 1, a transparent polystyrene film was printed, cured by irradiation with ultraviolet rays, a test piece was obtained, and a reflection density at various glucose concentrations was measured using a cyan light filter. Table 4 shows the measurement results.
なお,通常,マゼンタ(紅)系の色は,反射濃度の差が
それほど大きくなくとも,目視では十分区別できるもの
であり,実施例3においても,目視でも十分グルコース
濃度の差を判定できた。 Normally, the magenta (red) color can be visually distinguished sufficiently even if the difference in reflection density is not so large, and in Example 3 as well, the difference in glucose concentration could be sufficiently judged visually.
実施例4. 2−ヒドロキシ−2−メチルプロピオフェノンを用いな
かった以外は,実施例3と同様にして,印刷インキ組成
物Gを得た。得られた印刷インキ組成物Gを,実施例2
と同様にいて,基材に印刷し,電子線照射によって硬化
させ,試験片を得,実施例3と同様にして測定した。測
定した結果を表5に示す。Example 4. A printing ink composition G was obtained in the same manner as in Example 3 except that 2-hydroxy-2-methylpropiophenone was not used. The resulting printing ink composition G was used in Example 2
In the same manner as described in (1), a substrate was printed, cured by electron beam irradiation, and a test piece was obtained. Table 5 shows the measured results.
表4と表5との比較から,これらの場合には,電子線硬
化によって得られた試験片の方が,紫外線硬化によって
得られたそれより,発色の差が明確であることがわか
る。これは,電子線硬化と紫外線硬化とでは,得られる
硬化物の架橋度が異なるためと考えられる。 From the comparison between Table 4 and Table 5, it can be seen that in these cases, the difference in color development is clearer in the test piece obtained by electron beam curing than that obtained by ultraviolet curing. It is considered that this is because the degree of crosslinking of the obtained cured product differs between electron beam curing and ultraviolet curing.
また,電子線の照射線量7Mradを10Mradに代えて硬化さ
せると,発色は抑制されることもわかった。It was also found that when the irradiation dose of electron beam was changed from 7 Mrad to 10 Mrad and curing was performed, color development was suppressed.
比較例1. o−トルイジン 2.0部 イソブチレン−無水マレイン酸共重合体のブタノールエ
ステル「イソバン#10」 (クラレイソプリンケミカル(株)製,商品名) 2.5部 D,L−α−トコフェロール 0.1部 ポリオキシエチレンソルビタンモノオレート 1.2部 微結晶セルロース「アビセルSF」 (旭化成工業(株)製,商品名) 30.0部 n−ブタノール 47.0部 あらかじめ乳鉢で微粉砕したくえん酸無水物 3.2部 くえん酸ナトリウム無水物 12.0部 上記組成の混合物を十分にホモミキサーで微細分散させ
た後,グルコースオキシダーゼ(Grade II,東洋紡績
(株)製)0.5部およびパーオキシダーゼ(Grade III,
東洋紡績(株)製)0.1部を加え,さにホモミキサーで
微細分散させて,印刷インキ組成物Hを得た。得られた
印刷インキ組成物Hを実施例1と同様にして乾燥膜厚12
0μmとなるように透明ポリスチレンフィルムに印刷し,
65℃で30分間加熱乾燥し,実施例1と同様にして,試験
片を得,得られた試験片を種々の濃度のグルコース溶液
に浸漬したときの発色試験を行なった。反射濃度計を用
いて測定した平均の反射濃度は実施例1の場合とほぼ同
等であったが,グルコース濃度50mg/dlを超える領域で
は,点状の濃青緑色に発色する部分と発色のうすい部分
とがあらわれ,グルコース濃度が高くなるにつれこの傾
向が強くなることが観察され,目視判定が困難であっ
た。Comparative Example 1. o-Toluidine 2.0 parts Isobutylene-maleic anhydride butanol ester "isoban # 10" (Kuraray Isopurine Chemical Co., Ltd., trade name) 2.5 parts D, L-α-tocopherol 0.1 part poly Oxyethylene sorbitan monooleate 1.2 parts Microcrystalline cellulose "Avicel SF" (Asahi Kasei Kogyo KK, trade name) 30.0 parts n-Butanol 47.0 parts Citric acid anhydride 3.2 parts preliminarily finely ground in a mortar sodium citrate anhydrous 12.0 After thoroughly finely dispersing the above mixture with a homomixer, 0.5 parts of glucose oxidase (Grade II, Toyobo Co., Ltd.) and peroxidase (Grade III,
0.1 part of Toyobo Co., Ltd. was added and finely dispersed with a homomixer to obtain a printing ink composition H. The obtained printing ink composition H was dried in the same manner as in Example 1 to give a dry film thickness of 12
Print it on a transparent polystyrene film so that it becomes 0 μm,
A test piece was obtained by heating and drying at 65 ° C. for 30 minutes in the same manner as in Example 1, and a color development test was carried out when the obtained test piece was immersed in glucose solutions of various concentrations. The average reflection density measured using a reflection densitometer was almost the same as that in Example 1, but in the region where the glucose concentration exceeded 50 mg / dl, the spotted dark blue-green portion and the light-colored portion were formed. It was observed that some areas appeared and this tendency became stronger as the glucose concentration became higher, making visual judgment difficult.
比較例2. グルコースオキシダーゼ(Grade II,東洋紡績(株)
製)0.5部およびパーオキシダーゼ(Grade III,東洋紡
績(株)製)0.1部の代わりに,グルコースオキシダー
ゼ(タイプII,シグマ社製)0.2部およびパーオキシダー
ゼ(タイプII,シグマ社製)0.03部を用いた以外は,比
較例1と同様にいて,印刷インキ組成表Iを得,印刷
し,加熱乾燥し,試験片を得,得られた試験片を用いて
発色試験を行なったが,発色濃度が低く,目視でのグル
コース濃度の判定が困難であった。Comparative Example 2. Glucose oxidase (Grade II, Toyobo Co., Ltd.
0.5 parts and peroxidase (Grade III, manufactured by Toyobo Co., Ltd.) 0.1 part, glucose oxidase (type II, manufactured by Sigma) 0.2 parts and peroxidase (type II, manufactured by Sigma) 0.03 parts In the same manner as in Comparative Example 1 except that the printing ink composition table I was obtained, printing, heating and drying were carried out, a test piece was obtained, and a color development test was conducted using the obtained test piece. It was difficult to visually determine the glucose concentration.
本発明により,低温短時間で硬化乾燥できるために,検
査薬組成物中の酵素などの生物学的活性たんぱく質が失
活することなく,高価な生物学的活性たんぱく質を所要
量使用するだけで,簡便に,また,作業性および生産性
よく,体液成分検査用試験片を製造することができるよ
うになった。また,本発明により製造された体液成分検
査用試験片は,機器によるだけでなく,目視によって
も,正確な検査が可能なものである。According to the present invention, since curing and drying can be carried out at low temperature in a short time, biologically active proteins such as enzymes in the test drug composition are not inactivated, and only a required amount of expensive biologically active protein is used. It has become possible to easily manufacture test pieces for body fluid component inspection with good workability and productivity. Further, the test piece for inspecting a body fluid component manufactured according to the present invention can be accurately inspected not only by a device but also visually.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 正阿弥 重雄 東京都中央区京橋2丁目3番13号 東洋イ ンキ製造株式会社内 審査官 伊藤 明 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Shigeo Masaami 2-33 Kyobashi, Chuo-ku, Tokyo Toyo Inki Manufacturing Co., Ltd. Inspector Akira Ito
Claims (1)
薬および放射線硬化性化合物からなる印刷インキ組成物
を印刷し,印刷された印刷インキ組成物を活性放射線照
射により硬化することを特徴とする体液成分検査用試験
片の製造方法。1. A printing ink composition comprising a biologically active protein, an indicator and a radiation-curable compound is printed on a substrate, and the printed printing ink composition is cured by irradiation with actinic radiation. A method for producing a test piece for body fluid component inspection.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3280689A JPH06104073B2 (en) | 1989-02-14 | 1989-02-14 | Manufacturing method of test piece for body fluid component inspection |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3280689A JPH06104073B2 (en) | 1989-02-14 | 1989-02-14 | Manufacturing method of test piece for body fluid component inspection |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH02211897A JPH02211897A (en) | 1990-08-23 |
JPH06104073B2 true JPH06104073B2 (en) | 1994-12-21 |
Family
ID=12369079
Family Applications (1)
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---|---|---|---|
JP3280689A Expired - Lifetime JPH06104073B2 (en) | 1989-02-14 | 1989-02-14 | Manufacturing method of test piece for body fluid component inspection |
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JP (1) | JPH06104073B2 (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7156514B2 (en) * | 2004-04-30 | 2007-01-02 | Lexmark International, Inc. | Inks and printheads with internal clog prevention |
CN102326077A (en) * | 2009-02-18 | 2012-01-18 | 索尼公司 | The printing bioactive materials |
JP2013244611A (en) * | 2012-05-23 | 2013-12-09 | Dainippon Printing Co Ltd | Biomolecule ink printed matter, and manufacturing method therefor |
JP2016052799A (en) * | 2016-01-21 | 2016-04-14 | 大日本印刷株式会社 | Biomolecule print and producing method thereof |
-
1989
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