CN107884561A - 汗液模拟、收集和感测*** - Google Patents

汗液模拟、收集和感测*** Download PDF

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CN107884561A
CN107884561A CN201711171128.4A CN201711171128A CN107884561A CN 107884561 A CN107884561 A CN 107884561A CN 201711171128 A CN201711171128 A CN 201711171128A CN 107884561 A CN107884561 A CN 107884561A
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sweat
solute
biomarker
sensor
fluid
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乔舒亚·A·哈根
詹森·C·海肯费尔德
伊恩·帕帕茨克基
侯琳琳
拉杰什·奈克
南希·凯莉-洛克南
莫利·斯通
约翰·巴斯比
王霄
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University of Cincinnati
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Abstract

通过收集汗液并确定汗液中所选择的化学物质的浓度或含义来对在血液中可能发现的生物化学物质进行测量。汗液可利用基于时间的间隔收集器(10)来收集并利用外部设备来进行分析。它也可利用柔性的化学电容器(50)进行一次性收集，或利用化学场效应传感器98进行连续收集。

Description

汗液模拟、收集和感测***
关于联邦资助的研究或研发的声明
本发明至少部分地在来自美国政府的支持以及由美国空军研究实验室颁发的确定为SAPGrant号1008512的基金下进行。美国政府在本发明中享有一定的权利。
相关申请
本申请要求于2012年4月4日提交的美国序列号61/620,069的优先权，其公开内容以其全文通过引用并入本文。
背景技术
在血液中携带的生物标记物或化学物质可以提供重要信息，所述信息使得人们能够诊断疾病、健康状况、毒素、特性、以及其它生理属性，甚至在出现任何体征之前作出诊断。确定生物标记物的浓度提供对接受检验的患者健康状况的有利洞察，所述生物标记物诸如为电解质白细胞介素-1α、白细胞介素-1β、白细胞介素-6、肿瘤坏死因子α、白细胞介素-8、血管活性肠肽、神经肽Y、P物质、降钙素基因相关肽，食欲素-A，以及许多其它的生物标记物。例如，促炎和抗炎细胞因子以及诸如神经肽-y的神经递质与许多应用相关，所述应用包括心脏负荷测试、中风诊断、疲劳和创伤后应激性障碍。已经针对所有这些不同的生物标记物研发出了血液测试，但是血液测试需要抽取血液并随后进行分析。作为另一个实例，诸如钠和钾的电解质的直接测量提供对水合(hydration)的评估。这目前是使用抽血进行临床诊断。除了是侵入性的之外，许多生物标记物(诸如应激性)会受到抽血本身的影响。其它测试流体亦考虑用于确定血液中的生物标记物，例如唾液、尿液和呼吸都被考虑，但是不好使用且极易受到污染。利用可植入式传感器来分析血液价格昂贵且是侵入性的，且存在着重大风险。
汗液现在目前已知携带这些生物标记物。然而，针对生物标记物来分析汗液产生几个问题，这几个问题包括可如何收集、测试汗液，以及可从汗液获得的生物标记物的信息如何与血液***中的生物标记物相关联。
发明内容
本发明的前提是实现汗液能够有效地收集并在连续或不连续的基础上进行分析，以便有效地提供血液***中生物标记物浓度的指示。
更具体地，在一个实施例中，本发明提供了汗液收集***，其允许在离散的时间段内收集多个汗液样本以允许随时间的变化对生物标记物进行分析。此外，通过测量多个生物标记物(其一种或多种是参比(reference)生物标记物)，人们可以确立测试样本中生物标记物量的比率，以提供血液***中非参比生物标记物的浓度或含义(meaning)的可靠指示。
进一步地，根据本发明，可利用电化学生物测定装置来收集并分析汗液，所述电化学生物测定装置诸如化学场效应传感器(Chem-FET)、离子选择电极分析以及阻抗谱。这可与离子电渗电极(iontophoresis electrode)和诸如毛果芸香碱的胆碱能试剂相结合以便诱导非身体活动主体中的出汗，然后可利用电化学生物测定装置将汗液收集并进行分析。
在一个具体实施例中，所述电化学生物测定装置可作为贴片佩戴，其中在延长的时间段内收集数据并响应于诸如手机等的外部设备利用射频识别技术(RFID)或蓝牙技术以及许多其它技术来传送数据。
附图说明
我们通过下面的详细说明和附图可进一步理解本发明的目的和优点，其中：
图1是根据本发明的基于时间的汗液收集***的俯视平面图；
图2是图1的汗液收集***的局部剖切开的横截面视图；
图3A-3E是如图1所示的收集***的臂部部分的横截面视图，示出汗液在一段时间内通过收集臂进入到收集垫的行进情况，其中以点画阴影(stippled shading)示出汗液。
图4是本发明替代性实施例的分解视图。
图5是图1中所示装置的电路图。
图6是本发明第二替代性实施例的分解视图。
图7是在图6中所示装置中使用的汗液收集器的示意性俯视平面图。
图8是图6中所示的组装装置的局部剖切开的示意性横截面视图。
图9是图6中所示装置的电路图。
具体实施方式
根据本发明，利用汗液来以非侵入性的方式确定血液中生物标记物的浓度或含义。如图1中所示，允许按时间顺序存储多个汗液样本的汗液收集器10包括中央汗液收集构件12，其具有通到中央流体传输芯部16的汗液收集中央基部14，所述中央流体传输芯部16通到多个臂18。芯部16还通到顶部部分20(只在图2中示出)，其允许汗液蒸发或存储，因此其也提供连续的流体流。
汗液收集构件12可由将容易传输汗液的任何材料形成。通常，诸如纤维素网状物的纤维网状物特别适用于该应用中。其可用将促进流体流动的亲水性物质进行处理。然而，将容易允许水性流体流动的任何材料可用作汗液收集构件。
汗液收集构件12靠于汗液多孔粘合剂层22上，其允许收集器10被粘附到个体的皮肤23上。不可透过水的柔性薄膜24将基部12与臂18分离，所述柔性薄膜24具有与汗液收集构件12的中央芯部16相应的中央开口26。
径向延伸的臂18靠于塑料薄膜24的上表面上。每个臂通到闸门(gate)构件28，其依次径向向外延伸到桥接构件30，最后到达收集垫32。臂18、闸门构件28、桥接构件30和收集垫32都覆盖有蒸气多孔膜34(在图1中未示出，在图2在示出)，该膜34也具有允许芯部16延伸穿过的中央开口36，所述中央开口36通到上部流体传输盘20，所述盘20只允许流体蒸发和流动，其促进了汗液从皮肤到收集构件12的不断迁移。盘20可进一步涂覆有超吸收性水凝胶(未示出)以便进一步促进流体流动。备选地，所述盘20可覆盖有类似于蒸气多孔膜34的蒸气多孔膜。汗液收集器10的各个组件可以以有些甚至排除在外的多种方式布置，只要它们满足本发明的操作原理即可。此外，这种类型的汗液收集器10也可证实有利于简单地按时间顺序收集和储存其它流体，所述其它流体的范围从唾液到诸如在环境采样应用中河水等非生物流体。
如图3中所示，闸门28用于延缓流体流动到收集构件32。如下面所解释的那样，每个闸门与所有其它闸门略微不同。每个不同的闸门延缓流体流动持续不同的时间段，以使汗液在不同的时间下到达每个收集垫，从而如图所示那样按时间顺序提供在时间T1-T12下的离散的汗液样本，其中所示的12个时间仅是示例性的而并非限制性的。闸门28可由各种不同的材料来形成。适于这种应用的一种特定的材料是水溶性的聚合物薄膜构件，其抑制流体流动，但是在类似汗液的流体的存在情况下逐渐溶解。一旦溶解，其就允许汗液流动通过闸门以便填充桥接构件30和垫32。不同的闸门28可由厚度不同的水溶性薄膜形成或可在每个闸门中调节薄膜的聚合物组分以便创建不同的溶解速率。例如这可通过选择具有不同分子量的薄膜来完成。一种优选的材料是聚环氧乙烷。通过改变聚环氧乙烷的分子量可调节这些薄膜在水中的溶解度，其中较低分子量的薄膜更易于溶解。备选地，可调节薄膜厚度，其中更薄的薄膜比更厚的薄膜溶解地更快。当闸门28溶解时，汗液将流动通过闸门。闸门28可完全溶解，或者其可只变成为汗液可透过的，例如通过嵌入有聚环氧乙烷聚合物的布或纤维素纤维薄膜来实现。
闸门28依次与所谓的桥接构件30流体连通。桥接构件30是简单的流体导管，所述流体导管在它们变成为适当程度的不可透过流体流或溶质扩散之前只允许流体流动有限的时间段。这些流体导管30将允许汗液从闸门流到收集垫32。但是在一定体积的汗液流动通过桥接构件30之后，所述桥接构件30会溶解或变成为使进一步的流体流或溶质传输不可透过，中断流体路径，并防止进一步的汗液溶质(离子、分子、生物标记物)传输进或传输出收集垫32。从而每个收集垫32将在由闸门28允许的时间下接收汗液样本。当桥接构件30溶解时，其将垫32上的样本隔离，这样生物标记物信息不会与在稍后的收集时间段下的汗液混合、由其稀释、或失真。本发明包括实现以下的该相同的基本功能的所有方法：采样和分离汗液样本、将它们以正确保存它们的方式进行存储、以及以汗液样本代表在采样时间下或附近所排出的汗液的方式对它们采样和存储。因此汗液收集器10可利用替代性的材料，所述材料以提供类似结果的任何方式以物理或化学方式改变，包括但不限于桥接构件，其由暴露于汗液一段时间下交联的聚合物制成以防止流体流动；和/或包括但不限于闸门，其基于缓慢溶胀并移动成与汗液收集器的下一个流体组件流体连接的材料。
桥接构件可以是具有可接受的溶解率的任何水溶性构件。具体地，桥接构件可以是具有纤维表面的聚环氧乙烷丝线，所述纤维表面允许汗液以毛细迁移的方式通过聚乙烯丝线。一旦丝线溶解，流体将不再流动到收集垫32，因为在塑料薄膜24的上表面上将会有空隙，从而防止进一步的流体流动。
可针对各种不同的生物标记物来检测衬垫32。具体地，可希望分析一个或一个以上的参比标记物以便确定参比生物标记物的量，并将其与非参比测试的生物标记物进行比较，其中参比生物标记物的浓度通常是已知的。这允许人们利用参比生物标记物与所测试的生物标记物的比率来确定所测试的生物标记物的浓度，而无需知晓所测试的汗液体积。通常的参比标记物包括已知的方法，诸如用于确定电解质平衡的那些方法。在汗液中发现的一些生物标记物会由于酶或其它形式的分解或断裂、存储、保存、化学反应而迅速降解，或其它化学物质、材料或组分可被包括在垫32内或与垫32一起以保存由生物标记物所提供的所需信息。
在使用过程中，时间特定的汗液收集器10通过利用汗液多孔粘合剂22将收集器放置于皮肤上而施加到个体的皮肤上，其中汗液多孔粘合剂22接触皮肤。通常情况下，粘合剂层将具有保护性的剥离层(release layer)(未示出)，所述剥离层在使用前被去除。个体然后可继续进行日常活动或运动活动等。毛果芸香碱可施加到收集器所施加的区域以在汗液收集器10被施加之前或之后诱导汗液形成，或汗液可通过天然存在的方法或其它刺激形式来产生。多个汗液收集器10的组件可被堆叠并与其它桥接构件或闸门连接，以增加汗液收集器10的使用持续时间和总的样本收集。
在图3A-3E中示出为点画的汗液当产生时然后将沿着延伸到多个臂18的中央芯部16行进(参见图3A)然后到达各个闸门28(参见图3B)。第一闸门将在时间T1下溶解，以及第二闸门在时间T2下溶解，以及第三闸门在时间T3下溶解，依此类推。当闸门溶解(参见图3C)时，汗液可在溶解的聚环氧乙烷薄膜的表面上方迁移通过。聚环氧乙烷的纤维桥接构件30将允许汗液立即迁移到相应的收集垫32(参见图3D)。在经过一段时间之后，桥接构件30将溶解(参见图3E)，因为桥接构件狭窄且通过毛细管破裂而断裂，因此没有或没有足够的聚合物来允许汗液继续到达垫32。因此第一垫将在时间T1+X下停止收集汗液，以及第二垫在T2+X下停止收集汗液，以及第三垫在T3+X下停止收集汗液，依此类推，其中X是对于桥接构件30而言溶解所需的时间。时间X也可以是可控的变量，如果需要时间平均采样则尤其如此。在经过一段时间之后，收集器10被取出，且利用诸如质谱、比色光谱、电泳等标准分析和设备或由本领域内那些技术人员所公知的其它生物标记物的测定方法来针对某些生物分子对各个垫进行分析。这将提供基于时间的信息，其允许人们将在一段时间内的血流中的生物分子浓度与任何物理因素诸如应激性阶段相关联，或与药物的影响等有一定的关系。
如图4和图5中所示，本发明的替代性实施例是简称为一次性传感器50的一种一次性使用的汗液收集器/传感器50。该设备设计成在很短的时间段内快速地产生和吸收汗液，并允许即时电检测生物标记物。一次性传感器50全部是一次性的或者只是其部分是一次性的，其主要功能是以单个的(single)可穿戴的模式快速地刺激和感测汗液。一次性传感器50包括均固定在腕带58的化学电容器66或其它生物化学传感器以及盘状物54，所述盘状物用作诸如毛果芸香碱等胆碱能试剂的容器，其后跟随离子电渗电极56。如可以看出的那样，接线60从腕带通到控制一次性传感器50的第二设备。所述设备(未示出)例如可以是计算机、智能电话、或仅仅是设计成用于操作和分析从化学电容器66所提供的数据的专用装置。
化学电容器66被发现位于平面支撑层52上且部分地官能化有适于键联到被研究的一种或多种生物标记物上的一个或多个分子。这样，电容器66可以是一个非常简单的平面电容器或交叉指型电极，这取决于被检测的生物标记物的数目。支撑层52包括一系列的孔69，其允许毛果芸香碱移动通过层52以及允许汗液在相反的方向上流动到电容器66。也可包括或不包括参比电极、接地电极以及工作电极。其它合适的传感器包括化学场效应晶体管、化学电阻抗光谱仪以及电化学恒电位仪。
图5中示出适于装置50的通常接线图。在优选的实施例中，只有化学电容器66、盘状物54以及电极56实际上位于传感器的一次性部分上，而其余电路在分开的控制设备或外部装置(未示出)内。
在使用过程中，一次性的传感器50将用腕带58捆绑到个体手腕上。也可应用诸如粘合剂、胶带等的其它装置以便将传感器附接到身体部分。传感器的位置不是特别关键的，且可用在身体的具有足够表面积并接近汗毛孔的几乎任何部分。
一旦附接，接线60就附接到控制设备，以及离子电渗电极56被激活，导致毛果芸香碱从盘状物54迁移通过孔69。这将导致汗液的形成。由于化学电容器66与皮肤相邻，其会立即接触汗液。如果生物标记物存在于汗液中，这将通过电荷或电阻抗改变而立即检测到。通常情况下，多种生物标记物或化学物质将被检测到。具体地，一种生物标记物可以是参比生物标记物。剩余生物标记物的浓度或含义可基于参比生物标记物的浓度或含义来进行估计。一旦使用设定的时间段，则将传感器丢弃。基本上而言，可检测到存在于血液中的任何生物化学物质。对指示物理状态的主要生物标记物感兴趣。这些包括但不限于电解质、葡萄糖、乳酸盐、促炎细胞因子、抗炎细胞因子、儿茶酚胺、神经肽、以及可存在于血液中的任何蛋白质。
图6-9示出本发明的第三实施例。连续式传感器70包括汗液多孔粘合剂层72和汗液传输介质74。汗液传输介质包括靠于粘合剂层72上的收集垫76、通到蒸发或存储垫80的传输部分78。虽然以不同的阴影示出，但是收集垫76、传输部分78和蒸发垫80通常会由相同的材料制成。传输部分78从收集垫76延伸并围绕化学场效应传感器82。传感器70包括上部蒸汽多孔覆盖物84，其允许汗液蒸发，并由此继续流动通过收集垫。覆盖物84可包括水凝胶层(未示出)以便存储汗液，因此对汗液蒸发具有较小的依赖性。
汗液传输介质74设计成继续促进汗液仅仅在一个方向上传输通过收集垫76，否则收集垫会被填满且不能提供对新产生的汗液进行的连续的分析。因此，传输部分78从多个狭长的条带形成，其减少从收集垫76到蒸发垫80的流体流动路径。增大的蒸发垫80通过其尺寸并通过蒸发促进流体流动。从而允许通过传输部分78的减少的流动以及由蒸发垫80中的蒸发导致的流体流动的促进，提供汗液连续流动通过收集垫76到达蒸发垫80，而没有溶质或生物标记物的任何逆流或不希望的反向扩散水平。传输介质74可以是将吸收和传输水性流体的任何介质。其例如可以是任何织造或非织造网状物，以及特别是非织造网状物。纤维素纤维网状物特别适合用于这些用途。如果溶质或生物标记物随着时间的推移积聚或浓度过高，则蒸发垫80可被替换或用水清洗。蒸发垫可以是用于除去流体的任何材料或组分，包括水凝胶，其仅在它们吸收流体时通过芯吸将流体吸入且在尺寸上溶胀。因此蒸发垫可被更广泛地认为是一种简单的汗液去除元件。
化学场效应晶体管(Chem-FET)82包括由Kapton或类似的适当材料制成的柔性塑料基层92。射频识别(RFID)天线94和控制芯片96沿着基层92的周边93。依次地，化学型场效应晶体管82包括通过连接条带100连接到芯片96的传感器部分98。如图所示，在传感器部分98和射频识别(RFID)天线94之间的区域99是开放的，其允许毛果芸香碱定位在蒸发垫80或相邻的层(未示出)内以促进出汗。也可以包括与毛果芸香碱相邻的离子电渗电极。当组装时，传感器部分98直接靠于传输部分78上，当汗液从皮肤传输时对其进行分析。如箭头106所示，传输部分78和蒸发部分80位于传感器部分98的邻近层84侧上，以及收集垫76位于邻近粘合剂72的相反侧上。
优选地，传感器部分98是每个生物标记物具有三个以上相同化学场效应晶体管的栅极暴露的SiCMOS芯片。亚微米级的SiCMOS允许MHz阻抗谱。多步骤的图案形成/洗涤可用于固定生物识别元件。传感器在空间上被分成相同传感器的子组，或可利用诸如光引发化学图案形成技术来形成较大的传感器阵列。传感器允许持续地监测用于实现生物标记物特定传感器的较大阵列的多个生理状态。较大阵列可通过半特定的但不同的传感器通过统计确定来确定生理状态，而无需量化个体的生物标记物水平。
在另一个实施例中，传感器部分98是电极或电极阵列，其上涂覆有离子选择性材料。该离子选择性材料只允许一种类型的离子通过到达电极表面，从而允许单个分子类型的定量分析，诸如举两个例子而言的钠或钾。这些离子选择性传感器阵列能够确定个体的水合状态。
期望具有下述芯片96，其比印刷电路101稍厚，背离皮肤，但也优选在一个表面上形成所有器件。该化学场效应晶体管82通过电路电沉积到基层92的一个表面上而形成。然后将中央部分99切掉。尽管优选的是具有定位在传输部分78上的电路101，但是由于基层基板92的柔性性质，如果需要的话，传感器部分98可旋转180度，使得电路101直接抵靠收集垫76定位。
芯片96可被购买到。各种这样的芯片在市场上可商购到。可用于本发明的一种特定芯片是MLX 90129(由Melexis公司出售的)，在运行模式下其能够实现高达500微安培。该芯片具有内部温度传感器。芯片96也可以控制离子电渗电极(如果存在的话)。
在操作过程中，该装置将用粘合剂层72附接到皮肤。然后基于一定的周期例如每隔几分钟将诸如智能电话等阅读器紧密靠近传感器70捆绑，智能电话检测并记录诸如神经肽、细胞因子等所选择的生物分子的浓度、电解质平衡和体温。
连续式传感器70可以多种方式来改变以便提供额外的益处。例如，可以利用更强大的无线协议诸如蓝牙，或者可以使用替换性的通信或供电策略。例如，传感器可包括薄层电池，并提供其自身的电源，因此不依赖于RFID。RFID和蓝牙两者可以结合使用，其中当被提供适当的近场通信时，RFID可用于给电池充电。此外，可结合水凝胶的上层以便促进更大的汗液流动。其它的生物标记物检测方法和汗液传输方法可包括在内，只要它们提供连续或半连续地监测汗液中生物标记物的相同功能即可。
本发明的汗液收集器可提供各种不同的益处。基于时间的汗液收集器10可例如用在心脏负荷测试，允许心肌细胞因子生物标记物随时间变化进行映射，而无需血液导管。将毛果芸香碱以离子透入的方式(Iontophoretically)给药不到五分钟就可以刺激出汗以便使得这种测试持续。汗液收集器10然后可被定位且稍后利用诸如质谱等成熟技术进行分析。传感器10可被用作新药物治疗的按时间顺序的***反应的非侵入性的研究。其也可在适于改进的运动消耗或影响研究的定期运动活动中由运动员使用。
所述的一次性传感器50可用于针对非常特定的临界生物标记物进行快速地测试。例如，医护人员可将该设备通过将其捆绑到患者的手臂上而用在潜在的中风患者上，从而就没有必要寻找静脉。该设备将在2至3分钟内提供汗液，以及在稍后的1至2分钟内对生物标记物进行检测。三个主要的细胞因子，即肿瘤坏死因子、白细胞介素1以及白细胞介素6是在各种刺激诸如局部缺血后由培养的脑细胞所产生的。这提供将加快相应处理的诊断测试。通过选择合适的生物标记物，人们甚至可以区分缺血性与出血性中风。该非常特定的、廉价的传感器也可在各种不同的严格要求时间的测试中使用。
该连续监测，除了有利于对运动员进行基于时间的测试，且在临床研究中也可通过检测特定的细胞因子和神经肽被用于预防控制严重抑郁症的发作。它也可用于预测偏头痛，并且可用于连续地监测糖尿病。
一次性传感器和连续式监测器两者都可用于快速和/或连续地确定水合状态。
这是对本发明连同实施本发明的优选方法的描述，然而本发明本身应该只由所附的权利要求书来限定。

Claims (6)

1.确定汗液中第一溶质浓度的方法，包括：
收集一定体积的汗液；
确定所述汗液体积内的所述第一溶质的量，而无需确定所述汗液的体积；和
确定所述汗液体积中的第二溶质的量以便创建所述第一溶质与所述第二溶质的比率。
2.根据权利要求1所述的方法，其特征在于，所述第一和第二溶质选自于由下述构成的组：电解质、葡萄糖、乳酸盐、促炎细胞因子、抗炎细胞因子、儿茶酚胺、神经肽和蛋白质。
3.根据权利要求1所述的方法，其中所述第一溶质被用作参比生物标记物。
4.根据权利要求3所述的方法，还包括使用以下中的至少一个来确定个体的物理状态：所述第一溶质与所述第二溶质的比率；所述第一溶质和所述第二溶质的汗液浓度；和参比生物标记物的汗液浓度。
5.根据权利要求1所述的方法，其中确定三种或更多种溶质的汗液浓度。
6.根据权利要求5所述的方法，其中所述三种或更多种溶质中的一种溶质被用作参比生物标记物，所述方法还包括使用以下中的至少一种来确定个体的物理状态：三种或更多种溶质的汗液浓度；以及三种或更多种溶质中的至少一个比例；和参比生物标记物的汗液浓度。
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