CN1185478C - 无损伤性红外分光术中多光谱分析用的方法和装置 - Google Patents

无损伤性红外分光术中多光谱分析用的方法和装置 Download PDF

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CN1185478C
CN1185478C CNB971934010A CN97193401A CN1185478C CN 1185478 C CN1185478 C CN 1185478C CN B971934010 A CNB971934010 A CN B971934010A CN 97193401 A CN97193401 A CN 97193401A CN 1185478 C CN1185478 C CN 1185478C
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S·F·马林
G·哈利勒
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Abstract

本发明描述一种利用近红外和中红外区域内的多光谱分析确定试样中分析物浓度的方法和装置。在大约1100-5000nm范围内包含多个不同的非交叠区域的入射辐射被用来扫描试样。来自试样的漫反射辐射被检测，并且利用化学计量技术获得了指示分析物浓度的数值。各个非交叠波长区域获取的信息可以交差相关以去除背景干扰。

Description

无损伤性红外分光术中多光谱分析用的方法和装置
发明领域
本发明涉及利用多光谱分析确定试样中目标分析物浓度的方法和装置。本发明在化学分析中有广泛的应用，特别是在血液分析物的无损伤光谱光度分析中。
背景技术
血液中各种成份浓度的测定被广泛应用于人体疾病的诊断与治疗过程中。一种重要的应用就是血糖的测定。特别是，糖尿病患者应该定期监控其血糖浓度，对于胰岛素依赖型，即I型糖尿病患者更需要一天几次测定血糖浓度。另外，在防治冠状动脉疾病中测定的血液胆固醇浓度提供了重要的信息，并且在各种诊断过程中其它血液有机分析物(例如胆红素和酒精)的测定也是很重要的。
获取血液分析物浓度的最准确和常用方法是从病人身上提取血样，这些血样或者在实验室内采用高精度和高灵敏度的测定技术进行分析，或者采用精度较低的自检方法。特别是传统的血糖监控方法在每次测试时都需要从糖尿病人身上提取血样(例如利用指尖刺血针)并且采用血糖计(一种读取血糖浓度的分光光度计)或者比色计标度方法读取血糖水平。这种损伤性血液提取方式造成了糖尿病人的痛苦和负担并且由于所需测试次数较多增加了其感染的可能性。这些因素可能会导致糖尿病人在心理上对监控过程有抵触。
因此需要一种简单而准确的方法和装置以无损伤方式测定血液分析物浓度，特别是在糖尿病人血糖监控应用中。解决问题的一种途径是采用近红外分析的传统方法，其中利用某一或多个波长上测定的吸光度从给定试样中提取特定的分析信息。
液体试样的近红外吸收光谱包含了试样各种有机成份的大量信息。特别是与有机分子结构(例如碳-碳、碳-氢、碳-氮和氮-氢化学键)相关的振动、旋转和伸缩能量在近红外区域内产生可测的微扰量，它们与试样中的各种有机成份浓度有关。但是在复杂试样基质中，近红外光谱还包含了相当大的干扰，这些干扰的大小取决于分析物之间结构上的相似性、分析物的相对浓度水平、分析物之间的互扰关系以及特定系统固有的电学和化学“噪声”。这种干扰降低了利用近红外光谱测定法确定液体样本分析物浓度的测量效率和精度。尽管如此，人们仍然提出了一些以无损伤方式测定血液分析物的近红外装置和方法。
授予Purdy等人的美国专利No.5,306,004揭示了一种用于测定血液分析物浓度的方法和装置，其中采用具有不同的连续波长段的辐射照射人体。Purdy等人强调采用滤波技术来专门阻止波长位于水在近红外吸收光谱中两个波峰处的辐射透过，这两个波峰大约在1440和1935nm处。这种选择性阻挡的目的是避免人体中水份对光的吸收所引起的加热效应。
与此相反，授予Yang等人的美国专利No.5,267,152揭示了一种元损伤装置和技术，它仅仅利用包含近红外水吸收峰的红外光谱部分(例如“水透射窗口”，它包括1300-1900nm之间的波段)来测定血糖浓度。光学方式控制的辐射射向组织源并且随后由集光球聚集。聚集的辐射经过分析并利用存储的基准标度曲线计算血糖浓度。
人们还提出了用于测定复杂试样中分析物浓度的装置.
例如授予Richardson等人的美国专利No.5,242,602揭示了通过分析含水系统来检测多种活性或非活性水处理成份的方法。这些方法包括成份物在200-2500nm范围内吸收谱或发射谱的测定以及采用化学计量术算法来提取所获光谱数据段从而对多功能指示剂进行定量分析。
授予Nygaard等人的No.5,252,829揭示了一种采用红外衰减测量技术测定牛奶试样中尿素浓度的方法和装置。应用多变量技术是为了利用最小平方差算法、主分量回归、多重线性回归或人工神经网络学习算法确定已知成份物的光谱分布。通过解析阻挡所需分析物信号分量的贡献进行标度工作。Nygaard等人由此揭示出一种测量多分析物红外衰减和补偿背景分析物影响从而获取更精确测定结果的技术。
授予Ross等人的美国专利No.4,306,152揭示了一种光学流体分析仪，它从设计上将难以分析的浑浊试样或液体试样中背景吸收(即流体试样的总体或基本光吸收水平)对测量精度的影响降低到最小限度。该装置测量感兴趣试样成份物的特征光吸收处的光学信号和背景吸收波长附近的另一信号，并且将它们相减以抑制分析物信号中的背景分量。
利用上述方法和装置所获信息的精度受制于背景(即非分析物)和在近红外区域也具有系统吸收光谱的试样成份引起的光谱干扰。背景噪声水平呈现的是内在的系统限制，当分析物非常少时这种限制就特别突出。针对这种限制，人们试图通过各种手段提高信噪比，例如避免水吸收峰出现从而提高光照强度，减少光谱信息的分析量，或者采用基于背景吸收近似的减法或补偿技术。虽然这些技术为此作了某种改进，但是还是需要一种能够更精确地测定液体试样中分析物浓度的方法和装置，这在血糖监控应用中尤其如此。
发明内容
因此本发明的首要目标是满足上述需要，它所提供的方法可以测定包含各种背景基质和严重成份干扰的试样中分析物的浓度。该方法解决了试样中各种成份结构相似性、分析物浓度相对大小以及各种试样成份和仪器差异引起的光谱干扰等问题。
该方法通常包括以下步骤：(1)识别出近红外波段内几个波长不同的非交叠区域，这些区域与分析物浓度具有较高的相关度；(2)使包含这些波长区域的入射辐射照射在试样上以获取衰减的光谱作为表征试样成份的结果；(3)检测光谱衰减的辐射；(4)测定非交叠波长区域内某一波长处光谱衰减的辐射强度；以及(5)对测量进行相关分析以获取表示分析物浓度的数值。
在本发明的一个方面中，所提供方法的特征在于为获取分析物特定信息，对来自近红外和中红外区域的光谱数据进行了分析。因此该方法包括识别出近红外和中红外波段内几个明显不同的非交叠区域，通常在1100-5000nm的范围内，这些区域与选定分析物浓度具有较强的相关度或者提供了有关测量和仪器参数的信息。
在本发明的另一方面，所提供的方法包括以下步骤：(1)在近红外波段内选择几个与分析物浓度具有较高相关度的明显不同的非交叠波长区域；(2)利用包含选定光谱范围的红外辐射照射试样以获取光谱发生变化的辐射；(3)以光学方式滤光光谱发生变化的辐射以隔离或突出每个非交叠区域的辐射部分；(4)聚集并利用检测器测量被滤光辐射的强度；以及(5)通过对滤光的辐射应用确定的数学模型获取表示分析物浓度的数值。
本发明的另一目标是提供一种光谱光度测量装置，用来测定包含各种背景基质和严重成份干扰的试样中的分析物浓度。该装置在结构上包括一排能聚集和测定试样反射的衰减辐射的检测器。该装置被用于多光谱分析以获取包含特定分析物信号与涉及仪器背景噪声的信号的光谱信息以及干扰光谱信息。滤光单元上采用化学计量术技术以提高分析物特定信息与分析物浓度的相关度并且构筑能够测定分析物浓度值的系统算法。在本发明的一个方面中，采用衍射光栅系统来获取分析物特定光谱信息，这些信息由能够同时分析最多几百个数据点或波长的线性检测器阵列检测。
附图的简要说明
图1为按照本发明的装置示意图，它包含能够分析近红外和中红外波段的线性检测器阵列。
图2为按照本发明的另一装置的示意图。
图3为进行体内血糖耐量性研究时的时间扫描曲线。
图4示出了利用本发明方法的无损伤方式测定的血糖浓度结果的曲线图。
实施发明的方式
在详细论述本发明之前，应该理解的是本发明并不局限于所述装置或方法的特定部件。还应该理解的是这里所用的术语其目的仅仅在于描述特定的实施例而无限定作用。值得指出的是，说明书和所附权利要求书中所用的单数形式“一”或“这个”包含复数含义，除非在文中有特别说明。例如“一个分析物”具有分析物的混合物的含义，“一个光学传送元件”具有两个以上光学传送元件的含义，“一个用于反射透过辐射的装置”具有两个以上这种装置的含义，“一个波长”具有两个以上波长的含义，“一种化学计量术算法”具有两种以上算法的含义，等等。
在下面的说明书和权利要求中，涉及的术语具有如下的含义：
“化学计量术”涉及数学、统计和模式识别技术在化学分析中的应用。例如参见Brown等人在分析化学(1990)62：84-101上的文章。文中的化学计量术被用于采用先进的信号处理和标度技术的无损伤诊断仪器。信号处理被用来改善分析信号中物理意义信息的易用性。信号处理技术的实例包括傅立叶变换、一阶和二阶导数以及数值或自适应滤波。
在化学计量术中，“标度”指的是为定量化而对与化学浓度有关的测量数据所进行的处理。特别是利用化学计量术方法的统计标度可以用来从复杂的数据组中提取特定的信息。这些标度方法包括线性回归、多重线性回归、局部线性回归和主分量分析。在其它应用中，可以利用人工神经网络、基因算法和旋转主分量分析进行标度。
检测一种复杂化学基质中一种或多种组分的信息的仪器必然依赖于分析算法(如利用化学计量术导出的这些)，以便揭示属于一种或多种化学组分特有的信息。可以采用化学计量术将未知的与经过定标的标准和数据库进行比较，提供先进的分组分析形式以及从未知试样中提取能够被用作统计和数学模型中信息的特征。
“主要成分分析(PAC)”是一种在将化学计量术技术应用于复杂基质中化学分析物分光光谱测量中能够进行的数据缩减方法。采用PAC来降低大量的相互相关变量的维数同时保持一种成分区别于另一种成分的信息。将原始一组相互相关的变量(例如吸收光谱)本征矢量变换为数量很少的代表了原始组变量中大部分信息的一组非相关主要组分(PC)变量，利用这种本征矢量变换可以实现这一缩减。新的一组变量按照使其头几个变量保持所有原始变量中出现最多变化排序。见Jolliffe L.T.等人的《主要成分分析》Sprinter-Verlag，New York(1986)。更具体说，每个PC是所有原始测量变量的一种线性组合。第一个是在观测变量的最大方差方向上的矢量。将接下来的各个PC选作代表测量数据的最大偏差并与以前计算的PC正交。因此，各个PC按照重要性的降序排列。
术语“加权因子”包括部分最小平方回归和/或主要成分回归的加权系数、或从任何统计定标获得的能够被用于计算未知试样值(如分析物的浓度)的任何常数。“波长加权因子”是在构造能够从光谱数据中突出特定波长信息的光学滤光片装置中采用的加权常数的一个实施例。可以采用特定波长信息来确定与接受分析的试样(例如分析物的浓度)有关的所需值。波长加权因子可以体现为特定的滤光片密度(例如中性或特定波长)、滤光片厚度等，这些参数都能利用上述统计定标技术来确定。
实现波长加权因子的滤光装置可以用来选择突出与选定分析物浓度具有较高相关度的波长。“较高相关”或者“紧密相关”指的是特定波长的吸收光谱与特定分析物浓度之间的定量关系，其中两个变量具有0.9以上的相关系数(r)。
“中性密度滤光片”是指具有平坦吸收光谱的标准光学滤光片装置。可以采用中性密度滤光片与滤光片系统中的相关滤光片一起提供加衰减分析物在所选波长上吸收的权因子和进一步改善系统所提供的相关性的准确度。中性密度滤光片可以具有足以等量地衰减所关注范围中所有波长的辐射的吸收光谱。
正如文中所采用的，“含水媒体”包括由水组成或者含有水的任何基质。因此，含水媒体包括水为主要成分的媒体，即含量至少约50％，以及水为溶剂但是含量低于约50％。这里把含水媒体具体限定为包括哺乳动物组织面内。
术语“血液分析物”是指在近红外范围中吸收谱的血液组分，血液组分的测量对病人监测或保健保障中是有用的。
正如这里所采用的，术语“近红外”包括在约660nm至3500nm光谱范围，通常在1050至2850nm，更经常地在约1100至2500nm范围的辐射。
术语“中红外”包括在约3501nm-6000nm范围的辐射。
术语“背景吸收”是指被分析的含水试样的整个或基准光学吸收，所选组分在一个或多个特征波长上的吸收要偏离这一背景吸收，达到表示所选组分浓度的程度。当背景吸收的基准高于所选组分的特征吸收，以致在复杂的含水媒体中找到大量干扰组分时，要准确地测量在所关注组分特征波长上吸收的小幅度变化需要应用这里所述的化学计量术技术。在所关注组分的总浓度相对地低于含水媒体时，例如在测量血液中的分析物情况中尤其是这样。
一般方法
提供一种利用近红外和中红外辐射确定液体试样中分析物浓度的分光光度法。为了获得能够被用于以更高的准确度确定分析物浓度的一组测量结果，本发明的方法与以前的技术不同，它利用了近红外范围中所含的所有光谱信息。
该方法包括步骤：(1)选择几个不同的非重叠的近红外波长区，一般在1100-3000nm之间，或者从近红外范围和中红外范围内选择，一般在3501-5000nm之间，其中每个区域限定一个光谱范围；(2)利用包含所选光谱范围的近红外光照射试样，获得已经被衰减的光谱上产生变换的辐射；(3)收集并测量包含在每一个所选光谱范围内的一个或多个波长上光谱上经过衰减的辐射的强度；(4)对这些测量结果进行相关，获得表示分析物浓度的值。
利用这一方法获得的光谱信息可以结合一些数学变换，以获得更精确的分析物浓度的值。例如，可以采用标准统计技术，如部分最小平方(PLS)分析、或主要成分回归(PCR)分析，使特定波长上的辐射吸收与分析物的结构和浓度相关。例如，Geladi等人(1986)Analytica Chimica Acta  185：1-17描述了PLS技术。对于PCR技术的描述，可以参考Jolliffe L.T.Principal Component Analysis，Pprinter-Verlag，New York(1986)。
于是，在从机体组织试样确定血液中分析物浓度中，一种方法涉及从1100至3500nm近红外范围中选择三个不相重叠的波长区。比较好的但并非必需的是，第一波长区在约1100至1350nm之内，第二波长区在约1430至1450nm之内或者1930至1950nm之内，第三波长区在约2000至2500nm之内，这里，每个区域限定一个“光谱范围”。第一区域包含蛋白质和其它细胞成分显示主要光谱活性的波长，第二区域以水的吸收光谱为主，第三区域包含分析物有机分子显露明显的光谱活性的波长。在这些组分不为主要物质的区域中，它们对吸收光谱也产生作用。于是，从每个区域获得的光谱上经过衰减的辐射含有大量的相关信息，必须利用统计方法减少这些相关信息，获得分析物的特定信息。
本发明还涉及采用信号处理来改善分析信号中重要物理信息的可及性。因此可以对特定波长上获得的信号的强度值进行处理，降低仪器噪声的影响。然后，利用已知的统计技术对经过处理的信号作多变量分析。
数据缩减的PCA方法是本发明中实际采用的减少大量相关变量的维数同时保留一种成分区别于另一种成分的信息的较佳方法。将原始的一组相互相关的变量(例如吸收光谱)本征矢量变换为数量很少的代表了原始组变量中大部分信息的一组非相关主要组分(PC)变量，利用这种本征矢量变换可以进行数据缩减。新的一组变量按照使其头几个变量保持所有原始变量中出现最多变化排序。
通过相对吸收比平均值的正交旋转可以对主要成分矢量进行变换，获得一个已知波长和属于分析物的波长上的吸收比的相对值。通过对三个光谱区中每个光谱区获得的信息进行这样的分析，经过线性算法对主要成分矢量进行交叉相关，以及利用减法去除干扰分析物的影响，所获得的值能够被用于系统算法中，确定分析物的浓度。
采用多变量技术来提供每个光谱区中特定波长上的辐射强度与特定试样基质(如机体组织)中分析物浓度相关的模型。利用两组同时获得的示范测量结果构造该模型，第一组测量结果，“预计组”包括光谱数据，如所选波长上的辐射强度，第二组测量结果，“定标组”包括利用元损伤性取样技术已经确定的较高准确度的分析物浓度。在分析物浓度的一个量程上进行这种过程，提供一组定标数据和一组预计数据。
在定标组和预计组中所获得的测量结果接受多变量分析，例如利用市场上可得到的多变量模型开发软件程序提供初始模型。将初始模型运用到预计数据中，导出能够与损伤性技术获得值进行比较的分析物浓度。通过逐次进行上述步骤，发展为一个精炼的数学模型，能够被用于建立系统算法，用于对采用本发明方法获得的数据进行分析。
在本发明的实施中，还采用了不同的非重叠光谱区的非分析物的特定信息，例如对每次光谱扫描进行归一化，减去背景和基线干扰，或者提供用于检测非准确测量结果的信号值。
在确定机体组织试样中血液的分析物浓度时，在约1320-1340nm光谱范围中获取的测量结果提供高反射、未经衰减的信号，因为光谱区中不存在主要吸收波段。通过收集和测量该范围内的辐射强度，获得的值能够被用于估测用以照射试样的近红外光的实际强度。可以用该值对各次扫描进行归一化并校正光源强度的起伏，光源强度的起伏会影响利用本发明方法获得的分析物浓度值的准确度。
另外，在约1430-1450nm或约1930-1950nm光谱范围内获取的测量结果提供了基本非反射的经过高度衰减的信号，因为在水的近红外吸收光谱中在约144和1935nm处出现的两个主吸收峰之故。通过收集和测量以一个或两个光谱范围内的辐射强度，获得的值能够被用作估测不完全被被照试样吸收的近红外光的强度。利用该值可以从其它光谱区中获得的分析物特定信号中减去背景或基线信息和/或为检测不准确测量结果提供内部参考。为了校正镜面反射引起的垫底效应(随皮肤质地和年龄而不同)，可以从利用本发明方法获得的每个光谱测量结果中减去该值。
从第一光谱区(例如跨越约1320-1340nm的光谱范围)获得的基本未经衰减信号的测量结果和从第二光谱区(例如约1430-1450nm或1930-1950nm)获得的经过高度衰减信号的测量结果也可以用于将漫反射辐射与镜面反射辐射进行比较。如果两个光谱区中的信号具有相对可比的值，那么，用于照射细胞组织试样的大部分辐射看来是被皮肤表面反射的，因此未能穿透皮肤与血液中分析物相互作用。利用这一信息可以识别由于未能获得细胞组织的适当仪器扫描而引起的元效测量。
在本发明的一个方面中，所提供的确定试样内分析物浓度的方法利用了从几个在红外区的不同的非交叠波长区域和适于现场或家用的光学处理系统得到的无损伤性测量结果。该方法一般包括以下步骤：(1)从近红外范围(比较好的是1100-3000nm范围)或者从1100-3500nm的近红外范围和3501-5000nm的中红外范围选择几个不同的非重叠的近红外波长区，其中每个波长区限定一个光谱范围；(2)利用包含所选光谱范围的近红外光照射试样，获得光谱变化的辐射，即反射的辐射；(3)对光谱变化的辐射进行滤光以隔离或突出来自各非交叠区域的辐射部分；(4)利用检测器收集并测量经过滤光的辐射的强度；以及(4)对这些测量结果应用精确的数学模型以获得表示分析物浓度的值。数学模型可以包含利用化学计量技术获得的相关算法。
利用许多分光光度计配置都能够执行本发明的方法。现在参考图1，以标号10从总体上表示确定液体试样中分析物浓度的特定装置。该装置包括辐射源12，它提供约1100至5000nm范围中的多个不同的不相重叠的波长区。本领域人员熟知许多合适的辐射源，如射向干涉滤光片的白炽灯光源、经相关调制盘调制的卤素光源、激光光源、激光二极管阵列、或高速发光二极管(LED)阵列。在一个特定装置中，辐射源12提供三个不同波长区的辐射，具体说，近红外的第一波长区，通常约为1100至1350nm的范围；第二波长区，通常约为1930至1950nm的范围；和第三波长区，通常约为2000至3500nm的范围。
装置10还包括试样接口光学装置14，它将来自辐射源的入射辐射射入含有分析物的试样媒体16。在与试样媒体接触后，收集从试样媒体上以漫反射光出射的光谱发生变化的辐射并将其送至多阶滤光装置，一般用18表示。
在不同的配置中，可以将试样接口光学装置14设计成能够使装置10与媒体16紧密相接，例如通过将装置置于试样媒体上与之直接接触，由此将辐射源置于紧靠被分析试样的地方而进行光束发射。光束发射后，利用光敏装置，如光束会聚装置或光束偏转光学元件收集反射辐射。另一方面，试样接口光学装置14可以包括与装置耦合的光纤波导，从而能够将装置置于远处和操作。提供其它一些配置，其中采用单束光纤将辐射送至媒体和从媒体接收辐射。设置在单束光纤一端的光极将近红外辐射发射到试样媒体16中并接收通过单束光纤返回到装置10的光谱上发生变化的辐射。可以采用蓝宝石或高等级的石英作上述光纤波导中的光学元件，因为这些材料在近红外光谱范围中具有很好的透射特性。
仍然参考图1，从试样16出射的反射光线被送至多阶滤光片装置18。具体而言，光线被送至包含可调滤光片装置20的第一阶，该装置能够根据外部产生或者装置10已经产生的信号调节其吸收特性。可调滤光片装置通常包括筛选滤光片，如中性密度滤光片，可以调节其吸收特性，改变对外部信号或系统命令所表示辐射强度的衰减。可调滤光片20所提供的衰减度与所选的预定因子有关，以保证从可调滤光片出射的辐射将维持恒定值，而不管预先滤光的辐射的强度如何。
从可调滤光片装置20出来的经过衰减的辐射与主分析物滤光片22相接，其光学特性能够有选择地让辐射源12发射的各个不相重叠的波长区中一个或多个波长通过。将通过主分析物滤光片的波长选择为与分析物的浓度相关。
在装置10中配备一个第二滤光片装置24，它与主分析物滤光片2相关，使得有选择地通过主分析物滤光片的波长与第二滤光片装置相互作用，第二滤光片装置选定的吸收特性使得每个通过波长的强度被其衰减。例如可以利用化学计量术技术导出的一组独立的加权因子确定第二滤光片装置所提供的衰减。
在一种特定结构中，利用从含有分析物试样获得的原始光谱的部分最小平方或主要成分回归确定加权因子。利用能够透射至少1100至5000nm范围辐射的合适基底层能够构造第二滤光片装置24。基底层上通常镀有一层或多层本领域内常用的金属和/或氧化物，提供多种衰减滤光片密度。利用感光乳剂或本领域专业人员熟知的化学汽相淀积(CVD)技术可以将这种涂层加在基底上。在另一种装置中，第二滤光片装置为光学密度的光谱线与利用旋转主要成分或最小平方分析技术确定的加权因子成正比的照相掩膜。
经过第二滤光片装置衰减后，各个波长与检测装置26，如PbS检测器、砷化镓检测器等相接。在特定的装置结构中，如果需要获取整个约1100-5000nm范围的测量，可以采用一个或多个硒化铅(PbSe)检测器。
从第二滤光片装置出射的波长经检测装置26检测并转变为能够应用分析物特定算法确定分析物浓度的信号。具体说，利用模拟-数字转换器将从第二检测装置获得的信号转变为数字信号。将数字化信息提供给微处理器或其它电子存储装置，这里利用该数字化信息提供能够在显示装置上看到的和/或记录在输出记录器上的分析物浓度。
在另一种结构中，装置10包括衍射光栅系统和线性检测器阵列以代替多阶滤光片装置18。从试样16反射的光线可以送至选择通过分立波长的衍射光栅系统，其中通过的波长与分析物浓度特别相关。随后通过的波长与诸如硫化铅检测器阵列之类的线性检测器阵列相接。在对整个1100-5000nm范围内进行测量的特定应用中，可以采用商标为MULTIPLEXIR的产品(可Graseby Infrcred，Orlando.Fla.购得)由获得硒化铅线性检测器。
如上所述，线性检测器阵列聚集和测量衍射光栅系统传送的波长以提供可以应用分析物专门算法确定分析物浓度的信号。
利用装置10能够获得各种复杂媒体，如具有复杂光谱背景的含水媒体中的分析物浓度的测量结果。在一种应用中，可以采用该装置确定血液中分析物的浓度，尤其是血液中的有机分析物，如葡萄糖、尿素(BUN)、类脂物、胆红素和乙醇，但不限于这些。血液中的分析物可能存在于体外的试样媒体(例如血样)或者本装置也可测量组织中的血液分析物。然而，装置10特别适合用于现场(例如测量血液中的乙醇)或者家庭健康监测(例如确定血糖含量)。
现在参考图2，标号50总体表示测量试样中分析物浓度的另一种装置。该装置包括辐射源52，它提供约1100至5000nm范围中的多个不同非重叠波长区的辐射。装置50还包括试样接口光学装置54，它将辐射源的入射辐射与包含分析物的试样媒体56接触。在与试样媒体接触之后，从试样出射的光谱变化的辐射作为漫反射光线被聚集和送至通过特定波长光线的滤光片装置58。
在操作中，入射辐射经试样接口光学装置从辐射源52到达试样媒体，在一种构造下，试样接口光学装置可以设计成与得分析物试样紧密相接。在传送之后，利用光学活性装置(例如光聚焦装置，透镜或者光束偏转装置)聚集反射的光线。试样接口光学装置54可以包括与装置50耦合的光纤波导能够遥控装置放置和操作。如上所述，另一系统利用了单根光纤束向和从媒体后者辐射。
反射的辐射被导向包括多个用λ1、λ2、λ3、…λn表示的分立滤光片元件的滤光片装置58。滤光片装置58传递大部分被选定的波长范围，它们提供了特定分析物信息、测量背景信息和可以校正仪器变化或干扰效应的信息。选定的从滤光片装置出射的波长由包含用D1、D2、D3、…Dn表示的多个分立检测器单元的检测器60阵列检测。检测器排列成由单个分立的检测器检测从滤光片装置出射的每个选定波长范围。合适的检测器结构都是已知的并且可以包括例如Pbs和PbSe检测器阵列。每个检测器将检测的辐射转换为可以用来获取指示分析物浓度值的电学信号。
从检测器获得的信号可以利用模拟/数字转换器转换为数值信号，即指示检测波长强度的数字信号。数字化的信息随后输入微处理器作进一步的处理(例如应用系统算法)，或者信息可以经电子显示装置显示。从分立检测器获得的模拟信号与模拟/数字(A/D)转换器相连以转换为数字形式。模拟信号可以利用本领域熟知的技术在转换之前作前置放大。A/D转换器的数字信息随后输入微处理器以利用特定分析物的系统算法计算分析物浓度。微处理器通过对检测信号应用化学计量算法计算分析物浓度。利用递归标度法和统计模型技术(例如上述的化学计量方法)可以确定特定分析物算法。
在实施本发明中，滤光片装置58可以包括至少一个能够提高通过波长与分析物浓度相关度的吸收特性的分立滤光片元件。具体而言，滤光片装置可以包括一个或多个利用化学计量技术导出的独立加权因子组使通过波长的强度衰减的滤光片元件。这种加权因子可以通过对取自包含分析物的试样的原始光谱进行部分最小平方或主分量回归导出。
在另一结构中，滤光片装置58包括两阶滤光片，第一阶包括多个选择通过来自试样的衰减辐射的大部分波段。选择通过的波长包括分析物特定信息、与测量背景有关的信息和可以用来校正仪器变化或干扰效应的信息。滤光片的第二阶直接靠近第一阶，并且用来使来自第一阶的每个通过波长的强度衰减。两阶滤光片的第二阶可以是中性密度滤光片，它具有平坦吸收光谱，足以使滤光片第一阶的每个通过波长的强度衰减程度相等。
装置50可以用来确定各种复杂媒体中一个或多个感兴趣分析物浓度，例如在包含复杂光谱背景的含水媒体中。特别是本装置可以用于确定血液分析物的浓度，特别是诸如但是不局限于葡萄糖、尿素、胆红素、胆固醇和酒精之类的有机血液分析物。如上所述，可以利用体外试样分析血液分析物浓度，或者可以利用组织的近红外扫描进行分析，例如从前臂组织扫描获取的反射测量。
当装置50被用来从组织源获取血液分析物测量时，从光源52经试样接口光学装置54发射的入射辐射到达组织皮肤表面，例如到达前臂。试样接口光学装置使辐射以一定的角度射向组织从而可以被皮肤附近的组织物质吸收和漫反射。入射辐射的光谱由于血液和组织成份的红外吸收而发生变化。入射的近红外辐射部分被组织源内的血液成份吸收、扩散和反射。这种光谱变化的辐射包含了光学活性血液成份的特定信息。
在利用装置50确定血糖水平时，可以利用漫反射近红外辐射检测和测量血糖分子的振动。振动包括血糖分子的转动和平移，包括谐波振动和组合振动。这些振动中谐波振动是主要的并且大约在1670-1690nm的范围内。葡萄糖组合振动带大约在2120-2280nm范围内。葡萄糖在大约1320-1340nm的近红外范围内没有明显的光学活性。
因此装置50可以包括具有四个部分的滤光片装置58，第一部分使大约1300-1360nm范围的波长区域的反射辐射通过，第二部分使大约1430-1450nm或者1930-1950nm范围的波长区域的反射辐射通过，第三部分使大约1670-1690nm范围的波长区域的反射辐射通过，第四部分使大约2120-2280nm范围的波长区域的反射辐射通过。
滤光片装置第三和第四部分通过的波长的强度包含分析物特定信息。如上所述，第三和第四滤光片部分可以包括提高通过辐射与组织试样中葡萄糖浓度相关度的加权因子。从滤光片第一部分获得的信息可以用来估计每次测量时背景光谱的贡献，并且可以用来校正或归一化从第三和第四滤光片部分获取的测量。从第二滤光片部分获取的信号(水吸收信息)可以用作内部检验以识别无效的测量，例如因未能获得组织试样正确的仪器扫描而导致的测量，或者可以用来校正从第三和第四滤光片部分获取的测量中的温度变化。
应当明白，虽然结合具体的较佳实施例对本发明作了描述，但是，以上的描述以及下面给出的例子只是为了进行说明而非限制本发明的范围。对于本领域的专业人员而言，在本发明范围内的其它方面、优点和改进是显而易见的。
                             例子
利用本发明的方法能够获得无损伤性葡萄糖测量结果。具体说，在约1100nm至3500nm近红外光谱区中进行反射式光学测量。利用钨-汞(W-Hg)辐射源、硫化铅(PbS)检测器和扫描率为nm/0.4秒的仪器，从自愿者前臂收集光谱扫描。
找出许多特定的光谱范围作为能够用于从前臂组织扫描中确定葡萄糖浓度的信息。进行的体内葡萄糖耐量研究与无损伤性获得的体外血液中葡萄糖浓度确定相结合，确定特定的光谱区。具体说，图3示出在体内耐量研究期间获得的与次数有关的扫描。正如图中所能看到的，在研究的次数过程期间记录了约2120至2180nm范围上反射强度差明显变化。这种变化的增大与耐量试验期间血液中葡萄糖含量的增大直接有关，这表明2120至2180nm的范围含有葡萄糖的特定光谱信息。
一旦找出特定的光谱范围，利用四个不同光谱范围的信息可获得无损伤性葡萄糖测量结果。第一光谱范围包含在约1320至1340nm处产生的辐射。这个范围提供很强的反射信号，在这一范围中没有葡萄糖的主吸收波段。可以利用第一光谱范围上获得的信息对各次扫描进行归一化，以便校正辐射源的起伏以及由于机械扰动造成的变化。
第二光谱范围包含在约1440至1460nm或约1940至1960nm中任一范围上产生的辐射。由于是衰减漫反射辐射的水的强吸收波段，这些范围提供基本没有反射的信号。利用这些范围上获得的信息能够作为从其它测量中减去的背景和基准线。这些测量结果允许对镜面反射信号值引起的起伏作垫底调节，能够用作检测不适当的测量结果。
第三光谱范围包含在约1670至1690nm处产生的辐射。这个范围提供由于存在葡萄糖振动谐波波段引起的分析物的特定信息。
第四光谱范围包含在约2120至2280nm处产生的辐射。这个范围提供由于葡萄糖组合振动波段引起的分析物的特定信息。
利用第一范围获得的信号对其它光谱区的信号进行归一化。当对每次光谱扫描重复进行时，这一过程可消除与光源变化有关的问题并起提供内部参考的作用。于是极大地减小了由光学接口偏差(例如病人移动)引起的测量偏差。
从第三和第四分析物特定范围中获得的信号中减去第二范围中获得的信号，可以消除背景信息。用这种方式，校正镜面反射产生的垫底效应，这种效应会随皮肤质地和年龄而变化。
在分析物的化学计量术分析中可以应用第三和第四范围信号校正的归一化和基准线。图4示出第二与第三范围中信号之间的归一化之差。
正如从图4中所示结果中能够看到的，血液中葡萄糖含量的增大导致两个范围中信号差的增大。

Claims (26)

1.一种确定人体组织试样中有机血液分析物浓度的方法，包括以下步骤：
(a)在光路中利用波长在1100-5000nm范围内的入射辐射照射试样；
(b)收集从试样出射的反射辐射；
(c)用设置在光路中的检测装置接收从试样出射的反射辐射；
(d)将所检测的反射辐射转换为信号；
(e)分析所述信号以确定分析物浓度；以及
(f)在近红外吸收光谱中，尤其是水的近红外吸收光谱中，测量来自所述人体组织试样的辐射强度，以进行以下步骤中的至少一个：
提供参考以检测不准确的测量；以及
识别由于未能获得组织试样的适当仪器扫描而引起的无效测量。
2.如权利要求1所述的方法，其特征在于检测装置包含以下的其中一个：
硒化铅线性检测器阵列；
硫化铅线性检测器阵列；
从硒化铅、硫化铅和砷化镓中的任何一个构成的至少一个检测器。
3.如权利要求1所述的方法，其特征在于红外光谱包含波长在1100-1350nm范围内的光谱区域、波长在2000-3500nm范围内的另一光谱区域以及波长在以上两个范围之间的光谱区域。
4.如权利要求1所述的方法，其特征在于有机血液分析物选自葡萄糖、尿素、脂肪、胆红素和酒精。
5.如权利要求4所述的方法，其特征在于血液分析物为葡萄糖。
6.如权利要求1所述的方法，其特征在于所述近红外吸收光谱包括1430-1450nm和1930-1950nm的光谱范围。
7.如权利要求6所述的方法，其特征在于所述水的近红外吸收光谱包括1440nm和1935nm出的吸收峰值。
8.如权利要求1所述的方法，其特征在于来自所述人体组织试样的辐射强度包括从1320-1340nm的光谱范围中获得的实质上未衰减信号以及从1430-1450nm和1930-1950nm的光谱范围中获得的高度衰减的信号。
9.在确定人体组织试样中有机血液分析物浓度的方法中，一种识别无效测量的方法，包括以下步骤：
利用波长在1100-5000nm范围内的入射辐射照射试样；
收集从试样出射的反射辐射；
检测接收从试样出射的反射辐射；
将所检测的反射辐射转换为信号；以及
测量由波段中的所述信号所指示的镜面反射强度，其中实质上在所述波段中吸收所述辐射，而所述波段衰减漫反射辐射。
10.如权利要求9所述的方法，其特征在于所述波段是水的波段。
11.如权利要求9所述的方法，其特征在于所述波段包括1440-1460nm和1940-1980nm中的任何一个。
12.如权利要求9所述的方法，其特征在于还包括以下步骤：
将镜面反射测量与来自漫反射辐射实质上未衰减的波段的信号测量作比较；
其中实质上两个所述测量的可比的值指示无效测量。
13.一种确定人体组织试样中有机血液分析物浓度的装置，包括：
(a)将包含近红外和中红外光谱范围内多个不同不相重叠波长区的入射辐射照射试样的装置；
(b)收集从试样出射的反射辐射并将所述反射辐射射入光路的装置；
(c)设置在光路中的可调滤光片装置，这里，所述滤光片装置使光路中的辐射强度衰减；
(d)主分析物滤光片装置，它能够接收来自所述可调滤光片装置的衰减辐射并选择性地通过分立的波长，其中所述分立的波长与分析物浓度特别相关；
(e)第二滤光片装置，它能够接收主分析物滤光片装置出射的分立波长并使所述波长衰减；
(f)接收第二滤光片装置出射的衰减波长的检测装置；
(g)将被检测波长转换为代表所述波长强度的信号的装置。
14.如权利要求13所述的装置，其特征在于可调滤光片装置包含一中性密度滤光片，它与滤光系统中相关滤光片协同使用。
15.如权利要求13所述的装置，其特征在于第二滤光片装置包含与滤光片系统中相关滤光片一起使用的中性密度滤光片。
16.如权利要求15所述的装置，其特征在于利用加权因子建立第二滤光片装置提供的衰减。
17.如权利要求16所述的装置，其特征在于加权因子利用化学计量技术导出。
18.如权利要求17所述的装置，其特征在于加权因子利用分析物吸收光谱的旋转主分量分析导出。
19.如权利要求13所述的装置，其特征在于检测器装置包含硒化铅检测器。
20.一种确定人体组织试样中有机血液分析物浓度的装置，包括：
(a)将包含近红外和中红外光谱范围内多个不同不相重叠波长区的入射辐射照射试样的装置；
(b)收集从试样出射的反射辐射并将所述反射辐射射入光路的装置；
(c)设置在光路中的滤光片装置，所述滤光片装置包含具有第一阶和第二阶的两阶滤光片，第一阶包括多个片，配置为选择通过来自试样的至少一个波长的反射辐射，第二阶靠近第一阶并且能够使滤光片装置的第一阶出射的每一个选择通过的波长强度衰减，其中两阶滤光片装置的第二阶为中性密度滤光片；
(d)多个检测器，从而由分立的检测器检测滤光片装置出射的每个波长；以及
(e)将被检测波长转换为代表所述波长强度的信号的装置。
21.如权利要求20所述的装置，其特征在于第一阶包含多个分立的滤光片元件。
22.如权利要求21所述的装置，其特征在于至少一个分立的滤光片元件包含选择用来提高通过波长与分析物浓度之间的相关度的吸收特性。
23.如权利要求20所述的装置，其特征在于多个检测器包括硒化铅检测器。
24.如权利要求20所述的装置，其特征在于第一阶包含使范围大约在1300-1360nm的第一光谱分析区域的至少一个波长通过的第一部分、使范围大约在1670-1690nm的第二光谱分析区域的至少一个波长通过的第二部分、使范围大约在1930-1950nm的第三光谱分析区域的至少一个波长通过的第三部分和使范围大约在2120-2280nm的第四光谱分析区域的至少一个波长通过的第四部分。
25.一种确定人体组织试样中有机血液分析物浓度的装置，其特征在于它包括：
(a)将包含近红外和中红外光谱范围内多个不同不相重叠光谱区域的入射辐射照射在试样上的装置；
(b)收集从试样出射的反射辐射并将所述反射辐射射入光路的装置；
(c)设置在光路中的衍射光栅装置，其中所述衍射光栅装置配置为接收从试样出射的反射辐射并且选择性地通过分立的波长，所述分立波长与分析物浓度特别相关；
(d)一线性检测器阵列，以接收由衍射光栅装置出射的波长；以及
(e)将被检测波长转换为代表所述波长强度的信号的装置。
26.如权利要求25所述的装置，其特征在于线性检测器阵列包括硒化铅线性阵列检测器阵列。
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HUP9901855A2 (hu) 1999-09-28
BR9707245A (pt) 2001-09-11
CZ230498A3 (cs) 1999-07-14
TW459132B (en) 2001-10-11
US6236047B1 (en) 2001-05-22
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EP0877925B1 (en) 2003-07-16
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US6040578A (en) 2000-03-21
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ATE245279T1 (de) 2003-08-15
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