CN1985001A - 电化学分析装置和相关方法 - Google Patents
电化学分析装置和相关方法 Download PDFInfo
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- CN1985001A CN1985001A CNA2005800162607A CN200580016260A CN1985001A CN 1985001 A CN1985001 A CN 1985001A CN A2005800162607 A CNA2005800162607 A CN A2005800162607A CN 200580016260 A CN200580016260 A CN 200580016260A CN 1985001 A CN1985001 A CN 1985001A
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- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- Y10T156/00—Adhesive bonding and miscellaneous chemical manufacture
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- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T29/00—Metal working
- Y10T29/49—Method of mechanical manufacture
- Y10T29/49002—Electrical device making
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T29/00—Metal working
- Y10T29/49—Method of mechanical manufacture
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- Y10T29/49117—Conductor or circuit manufacturing
Abstract
提供了一种电化学检验装置,其具有基层和顶层,在基层上具有第一电极,在顶层上具有第二电极。两个电极被其中具有开口的间隔层分开,使得由顶面上的一个电极、底面上的另一个电极和由间隔中的开口的边缘形成的侧壁限定了接收样品的空间。将用于进行电化学反应的试剂沉积在一个电极上和接收样品的空间的侧壁上。
Description
本申请要求2004年5月21日提交的美国临时申请60/521,555的权益,其全部内容引入本文作为参考。
发明背景
本申请涉及用于检测样品中被分析物的存在或者数量的单一用途检验条形式的电化学分析装置和制造与使用上述装置的方法。
用于被分析物(如葡萄糖)的电化学检测的单一用途一次性检验条是已知的。在这些检验条中,样品被引入检验条以接触至少两个电极。被分析物的氧化或者还原以在两个电极间产生的电流的形式被观察到。例如,在导电池(cell)中使用葡萄糖检测,如图1所示,葡萄糖被酶(葡萄糖氧化酶)氧化而形成葡糖酸内酯和被还原的酶。酶的氧化形式的再生通过与被氧化的介质的反应而进行,结果产生被还原的介质。该被还原的介质将电子转移到一个电极,而在另一个电极处,电子被转移到被氧化的介质上,因此产生可观察到的电流。图2显示了使用酶/介质试剂系统在检验条中的作为时间函数的可观察到的电流。在此图中,t=0是样品施加的时间。如图所示,电流升高通过最大值,然后下降达到最终稳态的平台。在达到最大电流后,进行确定被分析物量的测量,并且通常是在达到稳态后进行。
在图2中,达到最大值之前,观察到可构成总测量时间重要部分的延迟。该延迟的持续时间取决于电极间的距离以及检验条中所用介质的流动性。介质流动性是介质本身的性质,即扩散系数,但是其也取决于其他的样品性质如血细胞比容和粘度。
为了使用户更方便,通常对被分析物检验条,特别地对葡萄糖检验条的改善集中在两个主要目标:较短的检验时间和较少的样品体积。在某种程度上,这两个目标已先后实现,因为较小的样品体积使用具有较小电极-间距的较小池,并且较小的电极-间距导致较短的反应时间。然而,这些池仍具有图2的电流/时间曲线,并且因此在能够进行测量前嵌入了延迟。本发明消除了这个延迟,并且因此完成检验所需的时间明显减少。
发明内容
根据本发明,提供了一种电化学检验装置,其具有基层和顶层,在基层上具有第一电极,在顶层上具有第二电极。两个电极被其中具有开口的间隔层分开,使得由顶面上的一个电极、底面上的另一个电极和由间隔中的开口的边缘形成的侧壁限定了接收样品的空间。
在导电池中,其中用于进行电化学反应的试剂沉积在一个电极上而非侧壁上,该装置产生如图2中所示的信号曲线。在本发明的装置中,试剂不仅沉积在该电极上,而且还沉积在接收样品的空间的侧壁上(图3)。这使得信号曲线没有滞后或者具有减少的滞后(图4)。尽管仅25%的侧壁高度由试剂涂布就能够实现明显的改善,但在优选实施方案中,试剂在侧壁全高度上铺开。
本发明还提供一种制造本发明检验条的方法。根据该方法,形成中间结构,其包括基层和置于基层上的间隔层。基层具有置于其上的第一电极,并且该电极通过间隔层内的开口而暴露。因此,第一电极/基层和开口间隔层的边缘限定了槽(well)或者沟(channel)。含试剂的液体被引入槽或者沟中,使得其至少部分、优选全部覆盖槽的侧壁。该液体然后被干燥,在槽/沟的底部(第一电极)和侧壁上留下试剂涂层。其后,在槽/沟上添加顶层和第二电极。
在该方法的优选实施方案中,间隔层在基层相反侧上具有粘合涂层和脱模薄片(release sheet),并且槽的侧壁通过脱模薄片向上延伸。试剂材料被引入槽/沟中,使得在干燥前用含试剂的液体覆盖至少一些部分的脱模薄片侧壁,并且优选在干燥后用试剂覆盖。随后除去脱模薄片而形成槽,其中侧壁基本上完全被干燥的试剂覆盖。
附图简述
图1显示用于葡萄糖检验条的基本化学反应。
图2显示了在试剂仅施加于槽底部的导电池检验条中作为时间函数的电流。
图3显示了根据本发明的装置的接收样品的空间的横截面。
图4显示了在根据本发明的检验条中作为时间函数的电流。
图5显示了根据本发明的装置的接收样品的空间的横截面。
图6显示了本发明方法的图解示意图。
图7A和B显示了本发明方法的图解示意图。
发明详述
本申请涉及通常用于血糖分析类型的电化学检验装置或者样条。
定义
如本申请说明书和权利要求书中所用的,术语“电化学检验装置”是指使用电化学分析对样品中的被分析物进行确定的装置,单独或者与可再用仪表结合。优选的电化学检验装置是通常已知用于家用确定葡萄糖水平类型的一次性单一用途的装置。
如本申请说明书和权利要求中所用的术语“被分析物”是指待被测量的样品的组分。具体被分析物的非限制性实例包括葡萄糖、血色素、胆固醇和维生素C。
如本申请说明书和权利要求中所用的术语“电极”是指电化学检验装置的组件,其将电子转移到物质中或者从物质中转移出电子,所述物质在被引入到所述装置的接收样品的空间内的样品中,并且该组件连接到或者可以连接到电路作为接触同一样品的电极间的电流或者电位差来确定存在着的电子转移的量。本发明装置的电极由导电材料制成,该导电材料与电化学池要来检测的具体被分析物一致。合适导电电极材料的具体实例包括金、碳、银、钯和铂。用于第一和第二电极的导电材料可以是相同的,或者它们可以是彼此不同的。在本发明的优选实施方案中,用于形成电极的导电材料是金。
如本申请说明书和权利要求中所用,术语“间隔”是指在装置的两个电极间提供电隔离的材料层。因此,间隔通常是绝缘材料,并且电极间的电接触仅仅在接收样品的空间内存在样品的条件下才发生。在优选实施方案中,间隔由绝缘材料的薄膜或者薄片形成。合适的材料的实例非限制性地包括聚酰亚胺、聚酯、聚对苯二甲酸乙二醇酯(PET)、聚碳酸酯、玻璃和玻璃纤维。间隔也可以通过绝缘层的沉积,例如通过在电阻涂层上喷涂而形成。开口可以在上述层中使用传统技术而形成,所述传统技术包括在限定的薄膜或者薄片中预切割开口、激光或化学蚀刻等。
如本申请说明书和权利要求中所用,术语“试剂”是指化学品或者化学品的混合物,当与样品混合时其允许电化学检验装置被用于确定样品中的被分析物。试剂不必足够多以进行这种确定,并且虽然不是优选的,但在引入到检验装置前,向样品中加入更多的化学品是可接受的。然而,试剂确实至少包含氧化还原活性物质,当使用该装置时,其在第一电极上被氧化并且在第二电极上被还原(或者反之亦然)。试剂可以包括多种作为电极间电荷载体的氧化还原活性物质。
如本申请说明书和权利要求中所用,短语“确定被分析物”是指并且包涵定性检测被分析物的存在(即被分析物是否以可检测量存在于样品中)、半定量检测(即被分析物的存在量是否大于预定的阈值)和定量评价(即确定存在的被分析物的实际数量)。
如本申请说明书和权利要求中所用,术语“覆盖”是指涂布所指定的表面。未要求完全覆盖,例如在小孔可能存在于涂层中的情况下,只仅仅要求覆盖剂分布在所指定的表面上。另外,没有排除涂层可能小于全部面积的情况,这是因为在具体装置中涂布方法的非故意缺陷。
如本申请说明书和权利要求中所用,短语“从被覆盖的电极连续延伸的侧壁部分”是指这样的涂层,其中在侧壁上在电极上的试剂涂层汇流到试剂涂层中。
本发明的装置
图3显示了根据本发明的装置的接收样品的空间的横截面。如图所示,底物层31具有置于其上的电极32。间隔层33在其中具有开口,其提供了侧壁34、34′。顶底物层35具有置于其上的电极36。接收样品的空间37由电极32和36以及侧壁34和34′界定,并且其包含干的试剂38。在可替代的实施方案中,接收样品的空间可以具有顶和底表面,其部分被电极覆盖并且部分暴露顶层。干的试剂38在接收样品的空间37的底面覆盖电极32,并且沿着侧壁34、34′向上延伸。
图4显示了根据本发明的装置的电流/时间曲线,其中干的试剂基本上覆盖了全部的侧壁。该图与图2比较显示了本发明的明显优点,即电流立即出现并且在更短的时间内实现稳态。
虽然无意束缚于任何具体机理,但相信这种效果的出现是因为电荷载体从开始就存在于接近两电极的位置,并且因此可以立即产生电流。相反,当将试剂单独施加于第一电极表面时,化学反应可能当样品添加时就立即开始,但是真正的电流直到介质(或者一些其他的氧化还原活性物质)从第一电极扩散到第二电极时才能流动。这花费了时间,并因此在观察到与被分析物有关的电流前存在延迟。另外,对于具有少量活性介质的试剂的情况,超过某一点时,反应甚至不能开始,直到逆反应存在。化学反应开始时的这种延迟是随着其他扩散过程所造成的延迟而累积的。
基于这种机理,理论预计时间上的延迟将与电荷载体必须移动以到达第二电极的距离的平方有关。这意味着,如果干的试剂和第二电极间的距离减小到1/2,则时间将减小到1/4,并且即使涂布25%的侧壁将导致达到电流最大值所需时间减小到大约1/2。因此,在本发明的装置中,用干的试剂涂布至少25%、优选至少50%、更优选至少75%并且最优选全部的延伸到第一电极之上的侧壁。
这种机理也清楚地说明在试剂中重要的组分是介质或者电荷载体。因此,如图5所示,在本发明的可替换实施方案中,将包含酶如葡萄糖氧化酶的试剂层51沉积在第一电极的表面上,并且沉积氧化还原活性涂层52以覆盖接收样品的空间的底部和至少部分侧面。
氧化还原活性涂层52、或者试剂涂层38可以包含装置中所用的氧化还原活性物质的氧化还原态。这可以是还原形式、氧化形式或者其混合形式。氧化还原活性物质的具体的非限制性实例是已知用于葡萄糖及其他介导的电化学检测系统中的氧化还原介质。如本申请说明书和权利要求中所用的术语“氧化还原介质”是指除了被分析物以外的化学物质,其在多步处理过程中被氧化和/或还原,使得将电子从被分析物转移到电化学池的电极中或者相反。介质的非限制性实例包括:
氰铁酸盐
[FeIII(CN)5(ImH)]2-
[FeIII(CN)5(Im)]3-
[RuIII(NH3)5(ImH)]3+
[RuIII(NH3)5(Im)]2+
[FeII(CN)5(ImH)]3-
[RuII(NH3)5(Im)H]2+
[(NC)5FeII(Im)RuIII(NH3)5]-
[(NC)5FeIII(Im)RuIII(NH3)5]0
[(NC)5FeII(Im)RuII(NH3)5]2-
二茂铁(Fc)和衍生物,包括但不局限于:
二茂铁单磺酸盐
二茂铁二磺酸盐
FcCO2H
FcCH2CO2H
FcCH:CHCO2H
Fc(CH2)3CO2H
Fc(CH2)4CO2H
FcCH2CH(NH2)CO2H
FcCH2SCH2CH(NH2)CO2H
FcCH2CONH2
Fc(CH2)2CONH2
Fc(CH2)3CONH2
Fc(CH2)4CONH2
FcOH
FcCH2OH
Fc(CH2)2OH
FcCH(Me)OH
FcCH2O(CH2)2OH
1,1′-Fc(CH2OH)2
1,2-Fc(CH2OH)2
FcNH2
FcCH2NH2
Fc(CH2)2NH2
Fc(CH2)3NH2
1,1′-Me2FcCH2NH2
FcCH2NMe2
(R)-FcCH(Me)NMe2
(S)-FcCH(Me)NMe2
1,2-Me3SiFcCH2NMe2
FcCH2NMe3
FcCH2NH(CH2)2NH2
1,1′-Me2FcCH(OH)CH2NH2
FcCH(OH)CH2NH2
FcCH:CHCH(OH)CH2NH2
Fc(CH2)2CH(OH)CH2NH2
FcCH2CH(NH2)CH2OH
FcCH2CH(CH2NH2)CH2OH
FcCH2NH(CH2)2OH
1,1′-Me2FcCHOCONHCH2
FcCH(OH)(CH2)2NH2
1,1′-Me2FcCH(OH)CH2NHAc
FcB(OH)3
FcC6H4OPO3Na2
三(菲咯啉)锇II和锇III(即Os-phen)络合物,包括但不局限于:
Os(4,7-dmphen)3
Os(3,4,7,8-tmphen)3
Os(5,6-dmphen)3
Os(bpy)3Cl2
Os(5-mphen)3
Os(5-Cl-phen)3
Os(5-NO2-phen)3
Os(5-phphen)3
Os(2,9-dm-4,7-dpphen)3
和同构的钌络合物,包括但不局限于:
Ru(4,7-dmphen)3
Ru(3,4,7,8-tmphen)3
Ru(5-mphen)3
Ru(5,6-dmphen)3
Ru(phen)3
[Ru(4,4′-diNH2-bipy)3]2+
三(联吡啶)锇II和锇III络合物(即Os(bpy)3),包括但不局限于:
Os(bpy)3
Os(dmbpy)3
和相关的钌络合物,例如:
Ru(bpy)3
Ru(4,4’-diNH2-bpy)3
Ru(4,4’-diCO2Etbpy)3
带有其他配位体的双(联吡啶)锇II和锇III(即Os(bpy)2)络合物,包括但不局限于:
Os(bpy)2dmbpy
Os(bpy)2(HIm)2
Os(bpy)2(2MeHIm)2
Os(bpy)2(4MeHIm)2
Os(dmbpy)2(HIm)2
Os(bpy)2Cl(HIm)
Os(bpy)2Cl(1-MeIm)
Os(dmbpy)2Cl(HIm)
Os(dmbpy)2Cl(1-MeIm)
和相关的钌络合物,例如:
Ru(bpy)2(5,5’diNH2-bpy)
Ru(bpy)2(5,5’diCO2Etbpy)
Ru(bpy)2(4,4’diCO2Etbpy)
其中,Et是乙基,bpy是联吡啶基,dmbpy是二甲基联吡啶基,MeIm是N-甲基咪唑,MeHIm是甲基咪唑,HIm是咪唑,phen是菲咯啉,mphen是甲基菲咯啉,dmphen是二甲基菲咯啉,tmphen是四甲基菲咯啉,dmdpphen是二甲基二苯基菲咯啉,phphen是苯基菲咯啉。另外,应该理解的是可以单独或者彼此结合地使用这些介质的还原或者氧化形式。
本发明的方法
本发明还提供了一种制造上述类型的电化学检验装置的方法。该方法在图6中示意性地举例说明。如图所示,间隔层61被置于第一电极62上。这可以使用绝缘膜或者薄片而容易地实现,所述绝缘膜或者薄片用粘合剂涂布在间隔层61的两面上。间隔层61具有开口63,通过这个开口使第一电极62暴露。这个开口可以是如图6所示的槽63的形式,或者是如图7A和B所示的沟73的形式。绝缘支撑物64在电极62之下。
包含氧化还原活性物质的液体试剂65被引入间隔层61中的开口63/73中,以这种方式覆盖至少一部分,但优选全部的被暴露的第一电极62和至少一部分开口63/73的侧壁66。在本发明一个实施方案中,这种结果是这样实现的:将开口63/73填充至足够深度以至少部分地覆盖侧壁。该结果还可以这样实现:随着分配管嘴上的移动液滴,沿着电极和侧壁施加试剂,使得其留下润湿的痕迹,或者在轨迹上使用喷墨或者相似的分配器,实现润湿的侧壁以及润湿的电极。应该注意到的是,在通常用于葡萄糖检验装置的小规模中,表面张力牵拉试剂以覆盖全部润湿的表面,使得试剂表面积最小化。这有助于试剂铺展到角落和向上壁上,如果已经润湿的话。
液体试剂65然后在开口63/73中干燥以形成干的试剂,其被置于覆盖第一电极62和至少一部分侧壁66的层中。干燥可以这样实现:简单地使该结构在空气中干燥、在所施加的空气流中干燥、加热、在被加热的空气流中干燥、在真空中干燥、或者在受热的真空中干燥。然后在间隔薄片的顶部上施加具有导电电极表面68的薄片材料67以形成第二电极,其横跨开口63面朝第一电极62。至接收样品的空间的进样开口可以通过修整横向通过开口63/73而这样形成的中间结构来产生,并且与电极一起形成接触而连接到外部仪表。形成装置的优选方法描述于美国临时专利60/521,555和美国专利申请10/908,656(2005年5月20日提交),其引入本文作为参考。
实施例
使用面向金电极和包含葡萄糖氧化酶、氰铁酸盐、缓冲盐和可溶性稳定剂的试剂来构造两个装置。在根据本发明的装置中,试剂沿侧壁向上延伸入第二电极附近。在对比装置中,试剂仅被置于第一电极上。当样品一进入检验条(t=0)时,就施加300mV电压(正极是具有试剂的第一电极;负极是不带试剂的第二电极)至各个装置。如图2所示,在对比装置中,最初很短的时间没有真实的电流,在大约1秒处电流才开始增加。电流未能显著增加,直到试剂能够溶解并且扩散到第二电极时,由此提供了允许电流流动的逆反应。图4显示了根据本发明的装置的电流曲线图。在这种情况下,几乎当样品一进入检验条时试剂就溶解并扩散,并且电流几乎立即开始流动(t=0)。因此,在大约3秒,在该装置的夹层几何电极结构中实现了稳态电流,相反,对于另一个装置为大约5秒。
Claims (17)
1.一种电化学检验装置,其包括:
底层底物、顶层底物和间隔,所述底层底物具有置于其上的第一电极,所述顶层底物具有置于其上的第二电极,所述间隔置于顶层底物和底层底物之间并且其中具有开口,由此限定了接收样品的空间,其具有底面、顶面和侧壁,所述底面具有置于其上的第一电极,所述顶面与第一表面相对并且具有置于其上的第二电极,且所述侧壁由间隔中的开口的边缘形成;和
包含氧化还原活性物质的试剂,当使用该装置时,其在第一电极处被氧化而在第二电极处被还原;
其中,在引入液体样品前,在检验装置中,试剂被置于覆盖至少一部分第一电极或第二电极和至少一部分侧壁的层中。
2.权利要求1的装置,其中试剂覆盖侧壁高度的至少25%。
3.权利要求1的装置,其中试剂覆盖侧壁高度的至少50%。
4.权利要求1的装置,其中试剂覆盖侧壁高度的至少75%。
5.权利要求1的装置,其中试剂覆盖侧壁高度的100%。
6.权利要求1至5中任一项的装置,其中氧化还原活性物质选自:
氰铁酸盐
[FeIII(CN)5(ImH)]2-
[FeIII(CN)5(Im)]3-
[RuIII(NH3)5(ImH)]3+
[RuIII(NH3)5(Im)]2+
[FeII(CN)5(ImH)]3-
[RuII(NH3)5(Im)H]2+
[(NC)5FeII(Im)RuIII(NH3)5]-
[(NC)5FeIII(Im)RuIII(NH3)5]0
[(NC)5FeII(Im)RuII(NH3)5]2-
二茂铁(Fc)
二茂铁单磺酸盐
二茂铁二磺酸盐
FcCO2H
FcCH2CO2H
FcCH:CHCO2H
Fc(CH2)3CO2H
Fc(CH2)4CO2H
FcCH2CH(NH2)CO2H
FcCH2SCH2CH(NH2)CO2H
FcCH2CONH2
Fc(CH2)2CONH2
Fc(CH2)3CONH2
Fc(CH2)4CONH2
FcOH
FcCH2OH
Fc(CH2)2OH
FcCH(Me)OH
FcCH2O(CH2)2OH
1,1′-Fc(CH2OH)2
1,2-Fc(CH2OH)2
FcNH2
FcCH2NH2
Fc(CH2)2NH2
Fc(CH2)3NH2
1,1′-Me2FcCH2NH2
FcCH2NMe2
(R)-FcCH(Me)NMe2
(S)-FcCH(Me)NMe2
1,2-Me3SiFcCH2NMe2
FcCH2NMe3
FcCH2NH(CH2)2NH2
1,1′-Me2FcCH(OH)CH2NH2
FcCH(OH)CH2NH2
FcCH:CHCH(OH)CH2NH2
Fc(CH2)2CH(OH)CH2NH2
FcCH2CH(NH2)CH2OH
FcCH2CH(CH2NH2)CH2OH
FcCH2NH(CH2)2OH
1,1′-Me2FcCHOCONHCH2
FcCH(OH)(CH2)2NH2
1,1′-Me2FcCH(OH)CH2NHAc
FcB(OH)3
FcC6H4OPO3Na2
Os(4,7-dmphen)3
Os(3,4,7,8-tmphen)3
Os(5,6-dmphen)3
Os(bpy)3Cl2
Os(5-mphen)3
Os(5-Cl-phen)3
Os(5-NO2-phen)3
Os(5-phphen)3
Os(2,9-dm4,7-dpphen)3
Ru(4,7-dmphen)3
Ru(3,4,7,8-tmphen)3
Ru(5-mphen)3
Ru(5,6-dmphen)3
Ru(phen)3
[Ru(4,4′-diNH2-bipy)3]2+
Os(bpy)3
Os(dmbpy)3
Ru(bpy)3
Ru(4,4’-diNH2-bpy)3
Ru(4,4’-diCO2Etbpy)3
Os(bpy)2dmbpy
Os(bpy)2(HIm)2
Os(bpy)2(2MeHIm)2
Os(bpy)2(4MeHIm)2
Os(dmbpy)2(HIm)2
Os(bpy)2Cl(HIm)
Os(bpy)2Cl(1-MeIm)
Os(dmbpy)2Cl(HIm)
Os(dmbpy)2Cl(1-MeIm)
Ru(bpy)2(5,5’diNH2-bpy)
Ru(bpy)2(5,5’diCO2Etbpy)
Ru(bpy)2(4,4’diCO2Etbpy)
或者其互补的氧化还原形式(被氧化或者被还原)。
7.权利要求1至6中任一项的装置,其中试剂还包括葡萄糖氧化酶。
8.一种制造电化学检验装置的方法,包括以下步骤:
(a)形成底层底物,其具有置于其上的第一电极;
(b)在底层底物上形成间隔层,所述间隔层具有形成于其中的开口,通过该开口使第一电极暴露,并且侧壁在开口内;
(c)将包含氧化还原活性物质的液体试剂引入到间隔层中的开口内,
(d)将液体试剂干燥而形成干的试剂,其中将液体试剂引入到开口中,以这种方式使得当干燥时形成干的试剂层,其覆盖至少一部分第一电极和至少一部分侧壁;和
(e)将顶层底物置于间隔层上,所述顶层底物具有置于其上的第二电极,所述间隔层被校准以使第二电极面对第一电极,由此形成接收样品的空间,其具有第一表面、第二表面和侧壁,所述第一表面具有置于其上的第一电极,所述第二表面与第一表面相对,具有置于其上的第二电极,所述侧壁由间隔中的开口的边缘形成。
9.权利要求8的方法,其中干的试剂覆盖侧壁高度的至少25%。
10.权利要求8的方法,其中干的试剂覆盖侧壁高度的至少50%。
11.权利要求8的方法,其中干的试剂覆盖侧壁高度的至少75%。
12.权利要求8的方法,其中试剂覆盖侧壁高度的100%。
13.权利要求8至12中任一项的方法,其中在步骤(b)中施加的间隔层包括粘合涂层和脱模薄片,其被置于朝远离第一电极的间隔层的一侧上,由此一部分侧壁由脱模薄片组成,该方法进一步包括在干燥液体试剂后除去脱模薄片以使粘合层暴露的步骤。
14.权利要求13的方法,其中将液体试剂引入到开口中,所引入的体积足以将开口填充至一定水平,使得至少部分地覆盖由脱模薄片形成的侧壁部分。
15.权利要求8至12中任一项的方法,其中引入液体试剂,所引入的体积足以将开口填充至一定水平,使得部分地覆盖开口的侧壁部分。
16.权利要求8至15中任一项的方法,其中开口是槽的形式,该槽由全部侧面上的侧壁所界定。
17.权利要求8至15中任一项的方法,其中开口是沟的形式,该沟仅由两个相对侧面上的侧壁所界定。
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