JP3810534B2 - Hematocrit value measuring element and method for measuring hematocrit value - Google Patents

Hematocrit value measuring element and method for measuring hematocrit value Download PDF

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Publication number
JP3810534B2
JP3810534B2 JP28540397A JP28540397A JP3810534B2 JP 3810534 B2 JP3810534 B2 JP 3810534B2 JP 28540397 A JP28540397 A JP 28540397A JP 28540397 A JP28540397 A JP 28540397A JP 3810534 B2 JP3810534 B2 JP 3810534B2
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Japan
Prior art keywords
hematocrit value
measuring
acid
reagent layer
blood
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Expired - Fee Related
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JP28540397A
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Japanese (ja)
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JPH11118794A (en
Inventor
佳子 宮本
信 池田
俊彦 吉岡
史朗 南海
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Panasonic Corp
Panasonic Holdings Corp
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Panasonic Corp
Matsushita Electric Industrial Co Ltd
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Priority to JP28540397A priority Critical patent/JP3810534B2/en
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Description

【0001】
【発明の属する技術分野】
本発明は、ヘマトクリット値測定用素子、および同素子を用いたヘマトクリット値の測定方法に関する。
【0002】
【従来の技術】
全血中のヘマトクリット値は、赤血球容積比と呼ばれ、貧血または多血症の診断に用いられる。現在、ヘマトクリット値の測定は、ガラス製の細管に血液を注入し、毎分12,000回転で4〜5分間遠心後、分離した血球部分の相対比を計測する方法が採られている。
また、赤血球数を自動的に測定する装置や、ヘモグロビン濃度を定量する試薬を用いて求めたヘモグロビン濃度から計算することも可能である。
【0003】
【発明が解決しようとする課題】
従来のヘマトクリット値の測定方法によると、高速遠心機が必要であり、場所を選ばずにどこでも簡便に測定を行えるものではなかった。
従って、本発明は、血液中のヘマクリット値を迅速かつ簡便に測定する素子および同素子を用いた測定方法を提供することを目的とする。
【0004】
【課題を解決するための手段】
上記の課題を解決するために本発明のヘマトクリット値測定用素子は、電気絶縁性の基板、前記基板上に形成された少なくとも二つの電極を含む電極系および試薬層を備え、前記試薬層が、酸解離指数pKが2〜11で常温において固体の酸を、供給される1滴程度の被測定試料に溶解したとき10〜100mMの濃度となる量含有することを特徴とする。
この測定用素子を用いてヘマトクリット値を測定するには、血液試料を前記試薬層に添加する工程および前記二つの電極間の電導度または電気抵抗を測定する。
【0005】
【発明の実施の形態】
図1は本発明のヘマトクリット値測定用素子の概略構成を示す平面図である。1はポリエチレンテレフタレートからなる電気絶縁性の基板を示す。この基板1上にスクリーン印刷により導電性ペーストを印刷してリード部分2、3と二つの電極部分4と5を形成してある。さらにスクリーン印刷により、リードを部分的に覆うように、電気絶縁性ペーストを印刷して電気絶縁層6を形成し、電極4及び5の露出部分の面積を一定にしている。
この基板上に、好ましくは電極4および5を覆うように試薬層7を形成する。
【0006】
基板上の試薬層7に血液試料が供給されると、試薬層中の酸が溶解し、試料液中の水素イオン濃度が増加する。しかしながら、血球中のヘモグロビン分子中に存在するイミダゾール基の強い緩衝作用により、イミダゾール環の窒素原子上に一部の水素イオンが捕捉される。血液試料の電導度は、血球量よりも溶存イオン濃度に支配されるため、前記の緩衝作用により血液試料の電導度は減少する。
一方、緩衝作用は、血球量すなわちヘマトクリット値の増加に伴い強くなることから、電導度の測定に基づいてヘマトクリット値を求めることができる。
【0007】
ここに用いる酸としては、平衡定数をKとしたときpK=ーlogKで定義される酸解離指数2〜11で、常温において固体の酸が適当である。また、試薬層に含有させる酸の量は、試薬層に供給される試料、通常血液の1滴程度に溶解して10〜100mMの濃度となる量とする。
本発明のヘマクリット測定用素子に用いる酸としては、以下の実施例に示すグルコン酸、クエン酸、2−(N−モルホリノ)エタンスルホン酸、およびリン酸二水素ナトリウムの他、リンゴ酸、シュウ酸、酒石酸などの脂肪族カルボン酸、あるいは安息香酸などの芳香族カルボン酸などを用いることができる。
【0008】
また、試薬層中には、親水性高分子を含有させることができる。親水性高分子は試薬層の溶解を促進するとともに酸の担体として働く。適当な親水性高分子としては、カルボキシメチルセルロース、ヒドロキシエチルセルロース、ヒドロキシプロピルセルロース、カルボキシエチルメチルセルロース、ポリビニルピロリドン、ポリビニルアルコール、ゼラチン及びその誘導体、アクリル酸及びその塩やメタクリル酸及びその塩の重合体、でんぷんおよびその誘導体、無水マレイン酸及びその塩の重合体から選ばれる一種類以上が用いられる。
【0009】
【実施例】
以下、本発明を実施例によりさらに詳しく説明する。
《実施例1》
図1のようにポリエチレンテレフタレートからなる電気絶縁性の基板1上に、リード部分2と3、電極部分4と5、および電気絶縁層6を形成した後、電極部分4および5を覆うように、グルコン酸20マイクロモルを水1ミリリットルに溶解した溶液を5マイクロリットル滴下し、50℃の温風乾燥機中で10分間乾燥させて試薬層7を形成した。
このような構成のヘマトクリット値測定用素子の試薬層7の部分に血液を供給した。ここに用いた血液のヘマトクリット値は25%、35%、45%、または55%のものである。血液供給から10秒後に電極4と5の間の電導度を測定したところ、ヘマトクリット値と電極間の抵抗値との間には図2に示すような相関関係があった。
【0010】
《実施例2》
実施例1と同様にして形成した電極部分に、クエン酸30マイクロモルを水1ミリリットルに溶解した溶液5マイクロリットルを滴下し、50℃の温風乾燥機中で10分間乾燥させて試薬層7を形成した。
実施例1と同様にして測定した抵抗値とヘマトクリット値との関係を図2に示す。
【0011】
《実施例3》
実施例1と同様にして形成した電極部分に、2−(N−モルホリノ)エタンスルホン酸10マイクロモルを水1ミリリットルに溶解した溶液5マイクロリットルを滴下し、50℃の温風乾燥機中で10分間乾燥させて試薬層7を形成した。実施例1と同様にして測定した抵抗値とヘマトクリット値との関係を図2に示す。
【0012】
《実施例4》
実施例1と同様にして形成した電極部分に、リン酸二水素ナトリウム30マイクロモルを水1ミリリットルに溶解した溶液5マイクロリットルを滴下し、50℃の温風乾燥機中で10分間乾燥させて試薬層7を形成した。
実施例1と同様にして測定した抵抗値とヘマトクリット値との関係を図2に示す。
【0013】
【発明の効果】
以上のように本発明によると、血液中のヘマトクリット値を簡便にかつ迅速に測定することができる。
【図面の簡単な説明】
【図1】本発明の一実施例におけるヘマトクリット値測定用素子の概略構成を示す平面図である。
【図2】実施例において測定された抵抗値とヘマトクリット値との関係を示す図である。
【符号の説明】
1 電気絶縁性の基板
2、3 リード
4、5 電極
6 電気絶縁層
7 試薬層
[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an element for measuring a hematocrit value, and a method for measuring a hematocrit value using the element.
[0002]
[Prior art]
The hematocrit value in whole blood is called the red blood cell volume ratio and is used to diagnose anemia or polycythemia. At present, the hematocrit value is measured by injecting blood into a glass tube, centrifuging at 12,000 revolutions per minute for 4 to 5 minutes, and measuring the relative ratio of the separated blood cell parts.
It is also possible to calculate from the hemoglobin concentration determined using an apparatus that automatically measures the number of red blood cells or a reagent that quantifies the hemoglobin concentration.
[0003]
[Problems to be solved by the invention]
According to the conventional method for measuring the hematocrit value, a high-speed centrifuge is required, and it has not been possible to easily measure anywhere regardless of location.
Accordingly, an object of the present invention is to provide an element for quickly and easily measuring a hematocrit value in blood and a measuring method using the element.
[0004]
[Means for Solving the Problems]
In order to solve the above problems, the hematocrit value measuring element of the present invention includes an electrically insulating substrate, an electrode system including at least two electrodes formed on the substrate, and a reagent layer, and the reagent layer includes: It is characterized by containing an acid dissociation index pK of 2 to 11 and solid acid at normal temperature in an amount of 10 to 100 mM when dissolved in about 1 drop of a sample to be measured.
In order to measure the hematocrit value using this measuring element, the step of adding a blood sample to the reagent layer and the electrical conductivity or electrical resistance between the two electrodes are measured.
[0005]
DETAILED DESCRIPTION OF THE INVENTION
FIG. 1 is a plan view showing a schematic configuration of a hematocrit value measuring element of the present invention. Reference numeral 1 denotes an electrically insulating substrate made of polyethylene terephthalate. A conductive paste is printed on the substrate 1 by screen printing to form lead portions 2 and 3 and two electrode portions 4 and 5. Further, an electric insulating paste 6 is printed by screen printing so as to partially cover the leads to form the electric insulating layer 6, and the areas of the exposed portions of the electrodes 4 and 5 are made constant.
A reagent layer 7 is preferably formed on this substrate so as to cover the electrodes 4 and 5.
[0006]
When the blood sample is supplied to the reagent layer 7 on the substrate, the acid in the reagent layer is dissolved, and the hydrogen ion concentration in the sample solution increases. However, due to the strong buffering action of the imidazole group present in the hemoglobin molecule in blood cells, some hydrogen ions are trapped on the nitrogen atom of the imidazole ring. Since the electrical conductivity of the blood sample is governed by the dissolved ion concentration rather than the blood cell volume, the electrical conductivity of the blood sample is reduced by the buffering action.
On the other hand, since the buffering action becomes stronger as the blood cell volume, that is, the hematocrit value increases, the hematocrit value can be obtained based on the measurement of conductivity.
[0007]
As the acid used here, an acid dissociation index of 2 to 11 defined by pK = −log K when the equilibrium constant is K and a solid acid at normal temperature is suitable. In addition, the amount of acid contained in the reagent layer is an amount that dissolves in about one drop of the sample supplied to the reagent layer, usually blood, to a concentration of 10 to 100 mM.
Examples of the acid used in the element for measuring hemagrit according to the present invention include gluconic acid, citric acid, 2- (N-morpholino) ethanesulfonic acid, and sodium dihydrogen phosphate shown in the following examples, as well as malic acid and oxalic acid. An aliphatic carboxylic acid such as tartaric acid or an aromatic carboxylic acid such as benzoic acid can be used.
[0008]
The reagent layer can contain a hydrophilic polymer. The hydrophilic polymer promotes dissolution of the reagent layer and serves as an acid carrier. Suitable hydrophilic polymers include carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxyethyl methyl cellulose, polyvinyl pyrrolidone, polyvinyl alcohol, gelatin and derivatives thereof, acrylic acid and its salts, methacrylic acid and its polymers, starch And one or more selected from polymers of maleic anhydride and salts thereof.
[0009]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples.
Example 1
After forming the lead portions 2 and 3, the electrode portions 4 and 5, and the electrical insulating layer 6 on the electrically insulating substrate 1 made of polyethylene terephthalate as shown in FIG. 1, so as to cover the electrode portions 4 and 5, 5 microliters of the solution which melt | dissolved 20 micromol of gluconic acids in 1 milliliter of water was dripped, and it was made to dry for 10 minutes in a 50 degreeC warm air dryer, and the reagent layer 7 was formed.
Blood was supplied to the reagent layer 7 of the hematocrit value measuring element having such a configuration. The hematocrit value of the blood used here is 25%, 35%, 45%, or 55%. When the electrical conductivity between the electrodes 4 and 5 was measured 10 seconds after the blood supply, there was a correlation as shown in FIG. 2 between the hematocrit value and the resistance value between the electrodes.
[0010]
Example 2
To the electrode part formed in the same manner as in Example 1, 5 microliters of a solution in which 30 micromoles of citric acid was dissolved in 1 milliliter of water was dropped, and dried in a hot air dryer at 50 ° C. for 10 minutes to form the reagent layer 7 Formed.
FIG. 2 shows the relationship between the resistance value measured in the same manner as in Example 1 and the hematocrit value.
[0011]
Example 3
To the electrode portion formed in the same manner as in Example 1, 5 microliters of a solution of 10 micromol of 2- (N-morpholino) ethanesulfonic acid dissolved in 1 milliliter of water was dropped, and in a hot air dryer at 50 ° C. The reagent layer 7 was formed by drying for 10 minutes. FIG. 2 shows the relationship between the resistance value measured in the same manner as in Example 1 and the hematocrit value.
[0012]
Example 4
5 microliters of the solution which melt | dissolved 30 micromoles sodium dihydrogenphosphate in 1 milliliter of water was dripped at the electrode part formed like Example 1, and it was made to dry for 10 minutes in a 50 degreeC warm air dryer. Reagent layer 7 was formed.
FIG. 2 shows the relationship between the resistance value measured in the same manner as in Example 1 and the hematocrit value.
[0013]
【The invention's effect】
As described above, according to the present invention, the hematocrit value in blood can be measured easily and rapidly.
[Brief description of the drawings]
FIG. 1 is a plan view showing a schematic configuration of a hematocrit value measuring element according to an embodiment of the present invention.
FIG. 2 is a diagram showing a relationship between a resistance value and a hematocrit value measured in Examples.
[Explanation of symbols]
1 Electrically Insulating Substrate 2, 3 Lead 4, 5 Electrode 6 Electrical Insulating Layer 7 Reagent Layer

Claims (2)

電気絶縁性の基板、前記基板上に形成された少なくとも二つの電極を含む電極系および試薬層を備え、前記試薬層が、酸解離指数pKが2〜11で常温において固体の酸を、供給される被測定試料に溶解したとき10〜100mMの濃度となる量含有することを特徴とするヘマトクリット値測定用素子。An electrical insulating substrate, an electrode system including at least two electrodes formed on the substrate, and a reagent layer are provided, and the reagent layer is supplied with an acid dissociation index pK of 2 to 11 and a solid acid at room temperature. An element for measuring a hematocrit value, which is contained in an amount of 10 to 100 mM when dissolved in a sample to be measured. 電気絶縁性の基板、前記基板上に形成された少なくとも二つの電極を含む電極系および常温において固体の酸を含む試薬層を備えるヘマトクリット値測定用素子を用い、血液試料を前記試薬層に添加して血液試料中に前記酸を10〜100mMの濃度に溶解させる工程、および前記二つの電極間の電導度または電気抵抗を測定する工程を有することを特徴とするヘマトクリット値の測定方法。Using a hematocrit value measuring element comprising an electrically insulating substrate, an electrode system including at least two electrodes formed on the substrate, and a reagent layer containing a solid acid at room temperature, a blood sample is added to the reagent layer A method of measuring a hematocrit value, comprising: dissolving the acid in a blood sample at a concentration of 10 to 100 mM; and measuring the conductivity or electrical resistance between the two electrodes.
JP28540397A 1997-10-17 1997-10-17 Hematocrit value measuring element and method for measuring hematocrit value Expired - Fee Related JP3810534B2 (en)

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