CN1591000A - 液体样品中物质浓度的测量方法 - Google Patents

液体样品中物质浓度的测量方法 Download PDF

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CN1591000A
CN1591000A CNA2004100789911A CN200410078991A CN1591000A CN 1591000 A CN1591000 A CN 1591000A CN A2004100789911 A CNA2004100789911 A CN A2004100789911A CN 200410078991 A CN200410078991 A CN 200410078991A CN 1591000 A CN1591000 A CN 1591000A
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O·W·H·达维斯
C·P·利奇
M·阿尔瓦雷茨-伊卡扎
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
    • C12Q1/006—Enzyme electrodes involving specific analytes or enzymes for glucose
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    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
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    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G01N33/5438—Electrodes

Abstract

一种测量例如血液或隙间流体一类液体样品中的葡萄糖之类的物质浓度的方法和装置。所用的测量装置具有一个工作传感器部分(6b)、第二个工作传感器部分(8b)和一个参考传感器部分(4b)。液体样品放入测量装置中，并测量在每一个工作传感器部分(6b、8b)上的与液体样品中的物质浓度成正比的电流。比较测得的电流建立差值。如果误差值大于预先确定的阈值，则给出误差指示；如果所述的参数差在预先确定的阈值内，则记录所述工作传感器部分的电流，作为有效结果。另外还说明了带有两个工作传感器(6b、8b)的一次性使用的测试带。

Description

液体样品中物质浓度的测量方法
本申请为中国专利申请号01808646.2的分案申请。
技术领域
本发明涉及测量液体中物质的浓度的装置，更具体地说但不是唯一地，涉及测量血液或隙间流体中的葡萄糖浓度的装置和方法。
背景技术
测量血液中葡萄糖值的装置对糖尿病患者是非常宝贵的，特别是患者自己可以使用的装置更是宝贵，因为他们可以监测自己的葡萄糖值，并服用相应剂量的胰岛素。因此，这种装置的精确性非常重要，因为读数不精确会导致服用错误剂量的胰岛素，这可能是非常有害的。
还有，在所有实际的血液中葡萄糖的测量系统中，至少装置的一部分，即与血液样品接触的部分，是一次性使用的。因为使用者一般需要大量的一次性使用零件，因此，降低一次性使用零件的成本就特别重要。
现在已知的葡萄糖测量装置更多地是使用电化学测量方法，而不是老式的比色测定法。一般的原理是测量分别称为工作传感器部分和参考传感器部分的两个传感器部分之间的电流。工作传感器部分包括上面涂有一层酶试剂层的电极，该试剂包括酶和一种电子媒质的化合物。当将一个电位加在传感器部分上时，由于电子通过酶从要测量的物质(酶基片)转移至电极表面而产生电流。所产生的电流与传感器部分的面积和测试样品中的葡萄糖浓度成正比。由于假设已知工作传感器部分的面积，因此，电流与葡萄糖浓度成正比。
技术上已发现，如果工作传感器部分不是完全被血液覆盖，由于传感器有效面积减小，因此得出不精确的结果。提出了解决这个问题的各种方法，其中二种方法公布在US 5628890和US 5582697中。这两种方法都是依靠血液在测试带的表面上的单向流动，并且两种方法都是通过检测在工作传感器部分下游的电极或传感器部分上有液体样品存在而开始测试测量的。
通过减小工作传感器部分的尺寸，当然可以减小液体样品不够和工作传感器部分不能完全被覆盖的问题。然而，工作传感器部分面积小使标定结果有更大的可变性。
本发明者发现，除了工作传感器部分覆盖不完全以外，不精确的结果也可由生产这种装置的测试带时偶然的缺陷和例如使用者对工作传感器部分意外的损坏造成。根据发明者所知，以前解决这个问题的唯一实际的方法是保证用于生产测试带的印刷过程尽可能的精确，并依靠适当的质量控制措施。
发明内容
本发明的目的是至少部分地克服上述缺点，并且当从本发明的第一个方面看时，要提供一种测量液体样品中物质浓度的方法。该方法包括：提供具有第一个工作传感器部分，与第一个工作传感器部分的面积相同的第二个工作传感器部分和一个参考传感器部分的测量装置；将液体样品装入所述测量装置中；测量在每一个工作传感器部分上产生与液体样品中的所述物质的浓度成正比的电荷载体的电流；比较以每一个工作传感器部分上测得的电流，建立参数差值；和如果所述参数差值大于预先确定的阈值，则给出误差指示；并且如果所述的参数差在预先确定的阈值内，则记录所述工作传感器部分的电流，作为有效结果。
根据本发明，有效地测量二次与物质浓度有关的电流，并将两次测量进行比较，其中每一次测量都可用来作为另一次测量的检查。
在电化学分析方面，本发明是特别有利的。这时，要测量浓度的物质例如血液中的葡萄糖与工作传感器部分上的元素例如酶试剂起反应，产生载体电荷，从而产生与液体中物质的浓度成正比的电流。
另外，本方法所用的测量装置是新颖的和具有创造性的。因此，从本发明的第二个方面来看，它提供了一种测量在液体样品中的物质浓度的装置，所述装置包括用于产生与液体样品中所述物质的浓度成正比的载体电荷的一个参考传感器部分和一个工作传感器部分，其中，所述装置还包括也是用于产生与液体样品中的所述物质和浓度成正比的载体电荷的第二个工作传感器部分，每一个所述传感器部分的面积相同。
可以看出，根据本发明，该测量装置对比由于产生电荷载体，而通过两个工作传感器部分的电流；并且如果两个电流大不相同，即一个传感器部分上的电流与另一个传感器部分上的电流相差太大，则给出误差指示。这种方法不但可以检测一个传感器部分是否被液体样品适当地覆盖，而且可以检测在每一个传感器部分中的制造缺陷，或者在制造以后，是否被损坏，因为在这种情况下，即使完全覆盖工作传感器部分，在受影响的传感器部分上将产生异常的电流。
根据本发明，唯一可以不需要考虑的缺陷或损坏的形式是，对两个工作传感器部分影响程度相同的缺陷或损坏。然而，与影响一个工作传感器部分的缺陷比较，从逻辑上说这是不太可能的，因此要对先前技术进行改进。实际上，这种相似性可以忽略不计。在任何情况下，本发明不是限于提供两个工作传感器部分。技术熟练的人可以选择三个或更多个工作传感器部分，以进一步减少所有这些工作传感器部分受到相同的缺陷影响的概率。
用另一种方式来看本发明时可看出，本发明提供了一种结构，对于工作传感器部分的给定的总面积和给定的样品最小容积，可通过将工作传感器部分的面积分成二个，可以检测工作传感器部分不适当的充满和缺陷或损坏的情况。
一些或全部传感器部分可以作为一个整体装置的一部分。但最好至少将工作传感器部分作在可除去的测验件上。因此，当从另一个方面看时，本发明提供了一种用于测量在液体样品中的物质浓度的测试件，它包括一个基底件；一个参考传感器部分；和两个设置在基底件上的工作传感器部分，每一个工作传感器部分使用中可产生与液体样品中所述物质浓度成正比的载体电荷，每一个所述工作传感器部分的面积相同。
最好，在基底件上设置一个参考传感器部分。
技术熟练的人知道，所提供的测量装置对于正确的使用是可以自动进行测试的，它对克服损坏和某些制造缺陷是有效的。特别有利的是，由于可以大量制造例如测量血液中葡萄糖值的测试带，因此该装置中的传感器部分设置在单独的测验件上。一般，这种测试带可被懒人使用，他不需要非常小心地对待这些带以防止损坏。因此，在优选实施例中，可除去的试验件包括一次性使用的测试带。
由于根据本发明，可以发现损坏或有缺陷的测试带而拒绝它，因此最终结果的精确性和使用者的安全不再只取决于高度的制造精度和正确小心的使用。至少在后一种情况下，本发明的优选实施例，与已知的结构比较，有一个附加的安全层。虽然不希望有大量的试验被拒绝，但在许多情况下，不给出不精确的结果更重要。
两个工作传感器部分在装置内可以方便地布置，或根据优选实施例，布置在试验件上。布置该装置或试验件时，应使液体样品在工作传感器部分上自由流动。最好是，液体样品受到约束，使它在工作传感器部分上单向流动。
最好，两个工作传感器部分，一个放在另一个的下游。这可以保证在另一个工作传感器部分开始被覆盖以前，一个传感器部分总是完全被覆盖，这样可以避免液体样品不够覆盖二个传感器部分，和每一个传感器部分只是部分地被覆盖相同的量的这种那怕是很小的可能性。然而，如果上述小的危险可以接受，根据本发明的结构可使放置传感器部分比已知装置有更大的灵活性，同时还可以防止使用容积不合适的液体样品或使用其他不正确的产品或产品损坏。最好，两个工作传感器部分位于参考传感器部分的下游。
由两个工作传感器部分产生的电流可能不能直接进行比较，这是因为传感器部分不相同。在这种情况下，测量装置最好对由一个或两个工作传感器部分得出的测量值进行加权，使该二个测量值标准化。这时，参数差可以是标准化的电流值之间的简单的算术差。最好，两个传感器部分包括相同的工作材料和具有相同的面积。最好，两个工作传感器部分基本上相同。这可以使得参数差直接就是传感器部分上相应电流之间的比较，以便确定物质浓度的测量是否可靠。
用于确定不精确测量的阈值可以适当地选择。一般，因为适当的值取决于测量过程固有的可变性，结果所希望的精度等，因此阈值可根据经验选择。在一定程度上，在因阈值设定得低而得出的精确度，与因太不精确而不予考虑的测量的比例之间进行折衷。阈值可以设定在这样的值上，即根据结果服用的胰岛素不会对病人造成严重的伤害。
参数差可以是一个绝对值，例如，在每一个传感器部分上测量的电流之差，但最好为无量纲的，例如一个或另一个测量电流的百分数。
虽然不是一定需要，最好是在经过预先确定的时间以后，测量电流。
用于计算物质浓度的实际电流值可以只是从一个工作传感器部分得到的值，但最好是电流值的综合-例如二个电流的和或平均值。这样做的优点是可以利用最大的有效工作面积，这有助提高所得结果的精确度。
本发明还提出一种确定施加到测试带上的电化学测量装置的液体样品的容积充分的方法，该方法包括以下步骤：
提供用于测量物质浓度的测量装置，该测量装置具有用于产生与液体样品中所述物质的浓度成正比的电荷载体的第一个工作传感器部分、用于也产生与液体样品中所述物质的浓度成正比的电荷载体的第二个工作传感器部分、以及作为第一和第二工作传感器部分的共用参考的参考传感器部分，其中第一和第二工作传感器部分和参考传感器部分是设置在一次性使用的测试带；
将液体样品施加到所述测量装置；
测量在每一个工作传感器部分处与液体样品中所述物质的浓度成正比的电流；
比较从每一个工作传感器部分上测得的电流，形成参数差；以及
当所述的参数差大于预先确定的阈值，建立一个误差条件，以指示样品的容积充分的缺少。
本发明的优选实施例为测量血液中葡萄糖浓度的装置，其中两个工作传感器部分和参考传感器部分设置在一次性使用的测试带上。
现用例子，参照附图来说明本发明的优选实施例。其中：
附图说明
图1表示根据本发明的测试带的基底件；
图2表示加在基底件上的碳轨迹的布局；
图3表示加在测试带上的绝缘层；
图4表示酶试剂层；
图5表示粘接剂层；
图6表示亲水性薄膜层；
图7表示测试带的覆盖层；
图8为没有使用根据本发明的方法得到的结果的图；和
图9为使用根据本发明的方法得出的，与图8相同的图。
具体实施方式
图1表示形成用于测量血液样品中的葡萄糖浓度的测试带的基底的一个长方形聚酯带2。图中孤立的表示一个基底件2，然而在实际中，在制造结束时，可从一张大的标准片材上切出一组这种测试带。
图2表示在这个例子中是用网板印刷加在基底件上的碳素墨水图形，但也可以使用技术上已知的任何适当的沉积方法。碳层包括4个互相电气绝缘的不同区域。在第一个轨迹4的远端形成参考/计数器传感器部分的电极4b。轨迹4沿长度方向延伸，在其近端形成一个连接终端4a。在第二和第三个轨迹6、8的远端，形成二个工作传感器部分的电极6b、8b；而在其近端，形成相应的连接终端6a、8a。第4个碳区域为一个连接桥10，它是在插入测试带时，使测量装置的电路封闭，以接通测量装置用的。
图3表示也可以用网板印刷法加上的下一个层。这是一个不溶于水的绝缘掩膜12，它在电极4b、6b、8b上形成一个窗口。该窗口控制露出的碳的尺寸，因而也是酶试剂层14(图4)与碳电极接触的尺寸。该窗口的尺寸和形状可使印刷在两个电极6b、8b上的酶斑点的面积完全相同。这表示，对于给足的电位，在有血液样品的情况下，通过每一个工作传感器部分的电流理论上相同。
酶层-在本实施例中为葡萄糖氧化酶试剂层14(图4)-印刷在掩膜12上，因而可通过掩膜上的窗口，印刷在电极4b、6b、8b上，分别形成参考/计数器传感器部分和二个工作传感器部分。然后，按照图5所示的图形，在测试带上印刷一个150微米厚的粘接剂层。为了清楚起见，与先前的图形比较，该图形放大了。三个分开的粘接剂区16a、16b、16c在一起，形成了它们之间的一个样品腔18。
在测试带的远端，层叠两块亲水性薄膜20(图6)，并用粘接剂16固定就位。第一块薄膜的作用是该样品腔18成为通过毛细管作用，可吸入液体和液体沿着它运动的一个细的通道。最后一个层表示在图7中，它是一个在远端有一个透明部分24的塑料保护覆盖带22。这可让使用者立刻知道是否使用了测试带，还可以粗略地用肉眼检查，血液加得是否够。
现在来说明测试带的使用。将测试带插入测量装置中。桥10使该装置中的电路完成闭合，因此可自动接通该装置。该装置有触点，与测试带上的终端4a、6a、8a连接。测量装置通过上述的终端，将400mv的电位加在计数器/参考传感器部分和两个工作传感器部分中的每一个部分之间。
然后将一滴血放在测试带的远端上。毛细管作用沿着上述样品腔18和在计数器/参考传感器部分和二个工作传感器部分上吸入血液。
经过预先确定的时间后，测量通过每一个工作传感器部分的电流和比较两个测量值。如果二个测量值的差大于10％，则在测量装置上显示误差信息，必需重复试验。然而，如果二个测量值彼此的差别在10％以内，则在该装置中将二个电流相加，并转换成葡萄糖值，显示在液晶显示器上。
为了举例说明根据本发明所得到的优点，利用上述这样制造的测试带进行比较试验。在试验中，将容积以幅度为0.2微升，从1微升增加至2微升的血滴，在保持葡萄糖浓度固定不变的条件下，加在测试带上。每一个容积重复8次试验。测量和记录在每一个工作传感器部分上测出的电流。结果表示在本说明书所附的表1中。
对于第一部分试验，将二个电流简单地加在一起，模拟具有综合面积的单一一个工作传感器部分。这些结果绘在图8中。
在试验的第二部分中，首先比较二个电流。只有当二个电流相差小于10％，才将它们加在一起，作为有效的结果。相差大于10％的值不予考虑。这个第二部分试验的结果绘在图9中。
立即可以看出，第二组结果精确得多，即它们变化小得多。另外，由于如果两个工作传感器部分完全被覆盖。则它们只给出彼此一致的结果，因此可以安全地假设，只有当二个工作传感器部分完全被覆盖时，实际上才给出结果，因此，第二组结果也比第一组结果精确得多。
这样，本发明的优选实施例可以检测和拒绝加在测试带上的样品不充分的那些试验，即测试带使用不正确的那些试验。同样，还可以检测和拒绝因损坏或制造缺陷而有缺陷的测验带。
在技术领域内，技术熟练的人会理解，在本发明的范围内，可以对上述说明的内容作许多变更。例如，本发明可用于测量任何液体中的任何适当物质的值，不仅仅是血液中的葡萄糖。另外，工作传感器部分不需要设在测试带上，而可以是整个装置的一部分。上述实施例中所用的10％的数值差纯粹是示例性的，可以选择任何适当的数值。
                                 表1
容积μL  工作电流1：μA     工作电流2：μA     差别％   检验到的误差     无误差检验
1  7.07     0.00     -706800     7.07
1 6.94 5.98 -16.2175732 12.92
1  5.53     0.01     -92050     5.54
1  6.99     7.09     1.42393909   14.09     14.09
1  7.34     7.02     -4.59016393   14.35     14.35
1  7.16     6.79     -5.49742078   13.94     13.94
1  7.01     3.47     -102.13441     10.48
1  7.07     5.69     -24.2578605     12.77
1.2  7.18     4.54     -58.2286847     11.72
1.2  7.00     6.78     -3.35055351   13.78     13.78
1.2  7.09     1.79     -297.032475     8.88
1.2  6.31     0.00     -157550     6.31
1.2  6.78     6.79     0.11788977   13.56     13.56
1.2  6.95     6.59     -5.4029443   13.53     13.53
1.2  6.62     6.28     -5.36795158   12.89     12.89
1.2  7.23     3.78     -91.2721502     11.01
1.4  7.16     6.90     -3.76811594   14.06     14.06
1.4  7.14     6.94     -2.88184438   14.08     14.08
1.4  7.17     7.02     -2.13675214   14.19     14.19
1.4  7.02     6.01     -1.5918958   13.93     13.93
1.4  6.95     6.91     -0.5788712   13.86     13.86
1.4  6.93     6.88     -0.72674419   13.81     13.81
1.4  7.09     6.92     -2.4566474   14.01     14.01
1.4  7.25     7.40     2.02702703   14.65     14.65
1.6  7.808     6.59     -18.4825493     14.40
1.6  6.774     6.589     -2.80770982   13.36     13.36
1.6  6.928     6.904     -0.34762457   13.83     13.83
 1.6     6.892     6.453     -6.80303735  13.35   13.35
 1.6     7.087     7.314     3.10363686  14.40   14.40
 1.6     7.257     6.947     -4.46235785  14.20   14.20
 1.6     6.501     6.306     -3.09229305  12.81   12.81
 1.6     6.811     6.755     -0.82901554  13.57   13.57
 1.8     7.145     6.536     -9.31762546  13.68   13.68
 1.8     7.021     6.612     -6.18572293  13.63   13.63
 1.8     6.917     6.828     -1.30345636  13.75   13.75
 1.8     6.971     6.78     -2.81710914  13.75   13.75
 1.8     7.016     6.941     -1.08053595  13.96   13.96
 1.8     6.977     7.179     2.81376236  14.16   14.16
 1.8     6.946     6.794     -2.23726828  13.74   13.74
 1.8     7.203     7.183     -0.27843519  14.39   14.39
 2     7.145     6.536     -9.31762546  13.68   13.68
 2     7.021     6.621     -6.18572293  13.63   13.63
 2     6.917     6.828     -1.30345636  13.75   13.75
 2     6.971     6.78     -2.81710914  13.75   13.75
 2     7.016     6.941     -1.08053595  13.96   13.96
 2     6.977     7.179     2.81376236  14.16   14.16
 2     6.946     6.794     -2.23726818  13.74   13.74
 2     7.203     7.183     -0.27843519  14.39   14.39

Claims (3)

1.一种确定施加到测试带上的电化学测量装置的液体样品的容积充分的方法，该方法包括以下步骤：
提供用于测量物质浓度的测量装置，该测量装置具有用于产生与液体样品中所述物质的浓度成正比的电荷载体的第一个工作传感器部分、用于也产生与液体样品中所述物质的浓度成正比的电荷载体的第二个工作传感器部分、以及作为第一和第二工作传感器部分的共用参考的参考传感器部分，其中第一和第二工作传感器部分和参考传感器部分是设置在一次性使用的测试带；
将液体样品施加到所述测量装置；
测量在每一个工作传感器部分处与液体样品中所述物质的浓度成正比的电流；
比较从每一个工作传感器部分上测得的电流，形成参数差；以及
当所述的参数差大于预先确定的阈值，建立一个误差条件，以指示样品的容积充分的缺少。
2.如权利要求1所述的方法，其特征为，包括在施加样品的一预先确定时间之后，测量每一个所述工作传感器部分的电流。
3.如权利要求1所述的方法，其特征为，所述被测量的物质为葡萄糖，每一个所述工作传感器部分产生与液体样品中的葡萄糖的浓度成正比的电荷载体。
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WO2001067099A1 (en) 2001-09-13
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PL364990A1 (en) 2004-12-27

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