基于差别度检验的电子舌应用方法学研究
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摘要
电子舌作为智能感官仪器,是随着感官品评技术的不断发展而发展起来的,并正在为人类发展发挥着重要作用。电子舌检测的突出特点是综合分析产品的性质,不需要对样品进行任何处理,是一种无损、快速的现代化检测仪器,而且与感官品评相比又具有客观、科学等特点,迎合了现代社会快速发展的需求。所以电子舌虽然只经历了不到三十年的发展,但是已表现出了巨大应用潜力,不仅实现了基本味物质的区分和辨识,而且渗透到了食品研究的各个领域。但是纵观电子舌应用的方方面面,基本是基于主成分分析统计方法进行区分和辨识,在表达样品差别程度方面研究甚少。电子舌的开发和利用的目的就是使感官品评工作科学化、客观化和量化以代替人的感官品评为终极目标。所以仅仅是对物质的区分是远远不够的,应该将样品间的差别达到一个量化的程度,这是今后电子舌取得广泛应用必须解决的问题。虽然也有研究者曾提出了用距离d来表征两类样品间相对差别,但是在很多方面还有待于深入研究。
本研究是基于智能感官领域相关研究存在的问题,着重从以下几个层面进行：(1)电子舌检测阈值和差别度理论研究,从仿生意义角度对经典心理物理学定律在电子舌物理传感器刺激响应方面的适用性进行研究,探讨基于差别度检验的电子舌检测理论依据；(2)规范出电子舌差别度检验的方法学框架,通过一定的手段将电子舌检测样品间的差别量化,建立样品间的差别度,科学的表征样品间的差别,并对电子舌检测不同样品的结果进行比较；(3)通过实例应用来对电子舌样品差别度表示方法的科学性进行验证；(4)对以电子舌和电子鼻为代表的智能感官仪器性能进行评价。本研究即从实际应用出发,解决电子舌应用中存在的实际问题。本课题主要研究结果如下：
1.定义了电子舌区分阈值的概念并规定了检测方法。差别阈为电子舌能够区分的样品的最小差别程度,即电子舌的灵敏度。将电子舌每个传感器每个频率段认为是一个单元,采用成对比较法,分别计算每个单元PCA区分的DI值,DI>0时是刚能区分开的临界值,而且是在高于该值时都能稳定的区分,所以认为当前的值是每个单元的差别阈值,再将各个单元综合起来,根据统计的每个单元的DI>0的数目与总的数目18的比值,为区分概率,即电子舌的区分能力Da%,当Da%达到50%或以上,所对应的浓度记为电子舌传感器阵列对该物质区分的差别阂值。根据上述方法分析和检测了10种味物质的差别阈值,并对电子舌与人的灵敏度进行了对比,通过比较可知,除了蔗糖、咖啡因和甘氨酸电子舌区分不如人灵敏外,对硫酸奎宁区分与人相近,其他的味物质如柠檬酸、酒石酸、AK糖、氯化钠、氯化钾、谷氨酸钠电子舌区分灵敏度要远高于人类。
2.定义了表征电子舌检测样品间差别度的概念、表示方法和差别度检验方法。
在几何空间上,对两个样品差别程程度大小的度量,即差别度(Differential degree, Dd),是通过计算两样品间的距离Da值来反映的,通过简单溶液分析,Dd值与物理刺激量呈现正相关,Dd值越大说明样品间的差别程度越大。Dd的引入将样品间不能度量的差别以具体数值的形式表现出来。建立了两种电子舌差别检验方法：一是成对比较检验法,确定一个对照样品R(性质稳定,且与所比较的样品属同一类)和一系列待检测样品(1,2,3,4……i),待检样品分别与对照样进行比较；二是顺序检测法,分别比较样品间的相对差别度,或将一系列样品分别与对照样进行比较。
3.通过对八种味物质溶液分析可知,电子舌检测样品的差别度Dd值与物理刺激强度间存在类似费希纳定律的现象。电子舌检测的样品Dd值与I(溶液浓度)间呈现对数关系,刺激强度按几何级数递增,而电子舌检测的样品间差别度按算术级数递增。电子舌检测样品间Dd值与I的关系与心理物理学研究中的费希纳定律类似,为今后电子舌的研究与应用奠定了一定的理论基础。
4.电子舌检测与感官品评技术结合实现了不同品牌绿茶饮料的区分,中国黄酒品牌区分、种类区分和不同年份区分,两种分析技术结果相互印证。
确定了茶味、酸度、甜度、苦涩感作为茶饮料感官品评的感官特性指标,确定醇香、酱香、酸、甜、苦、涩、辛辣感作为黄酒感官品评的感官特性指标。根据电子舌检测样品间的差别度Dd值大小可以判定样品间差别程度的大小。
对电子舌检测数数据采用DFA方法可以实现对不同品牌绿茶饮料进行辨识,辨识正确率100%；采用SIMCA方法可以实现绍兴黄酒与非绍兴黄酒的辨识,辨识正确率93.3%。
5.建立了产品货架期分析和质量控制与管理的电子舌差别检验和感官三点检验方法。
采用顺序检测法,分析了黄酒敞口放置时的质量变化情况。随着敞口放置时间的延长,Dd值有增大的趋势,即样品的差别度与时间具有正相关变化趋势。通过电子舌检测结合感官品评,12h可作为黄酒在室温下敝口放置的质量控制临界值,在12h或时间更长时,黄酒的感官质量明显变差,而且已经能够被人类所察觉。通过建立的Dd值与时间关系的数据模型可以对未知样品的质量和货架期进行分析预测,达到产品质量控制的日的。
采用成对比较法,分析橙汁饮料在不同条件处理时,根据Dd值判定其质量变化情况。窒温24℃处理对橙汁饮料的质量影响不大;而在高温37℃处理对其质量影响显著,Dd值随着处理时间的延长具有不断增大的趋势,将5h作为橙汁饮料在高温下质量控制临界点,当Dd值大于2.67时,产品就会发生较大的质量变化。
6.提出并构建了以检测效率、重复性、区分稳定性、灵敏性检测限这五个指标为标准的智能感官仪器(电子舌、电子鼻)性能评价方法,规范了操作方法和数据处理方法。依据构建的性能评价方法,对伏安电子舌的性能进行了研究；同时对本课题组开发的智鼻和某电子鼻的性能进行了研究和比较。
Electronic tongue (E-Tongue), one kind of intelligent sensory instruments, was produced with the development of sensory evaluation techniques, and in time, its applications have been increasing. This device is characterized by analysis of the overall characteristic, non-destructive process for the samples and designed for rapid detection. It has proven to be a good alternative for sensory evaluation techniques with objective and scientific, meets with the rapid development of the modern society. Less than30years since its development, it has been proven to be an ideal alternative for traditional chromatographic and in the analysis of food. Its great application potential has been expressed gradually; it has not only succeeded in distinguishing and identifying basic taste substances, but also in penetrating into various fields of food research. All of these studies have been mainly based on the principle component analysis (PCA) to distinguish and recognize substances qualitatively. Minimal research efforts in the quantitative expression of the sample difference have been undertaken. The purpose of the development and use of the E-Tongue is to make sensory evaluation scientific, objective, and quantitative, with the end view of using it in artificial sensory evaluation. Material distinction is not enough; the difference between samples must be quantified and resolved for a wider application of the E-Tongue in the future. Although researchers have proposed using a distance to characterize the relative difference between the two types of samples, and this research remains to be a concern worthy of further investigation.
This research was inspired by a related study in sensory field. In the intelligent sensory field, issues arise from the following aspects:first, the research of the threshold of the substances detected by E-Tongue, and the applicability of the classical law of psychophysics in E-Tongue and the exploration of the theoretical foundation based on difference test; second, the confirmation of the methodology used in the difference test for E-Tongue applications to qualify differences of the samples, establish the differential degree(Dd), characterize differences between samples scientifically, and compare the results of different samples; third, it is also the intent of the research to validate the science behind sample difference; forth, evaluate the performance of the intelligent instruments, such as E-Tongue and Electronic nose. This paper mainly embarked from the practical application angle, and resolved actual existing questions on the E-Tongue as a handy, rapid, and objective method. The results are showed as follows:
1. Established and confirmed the concept and test method of the threshold detected by E-Tongue. The difference threshold was defined as the smallest difference between samples distinguished by E-Tongue, and that was sensitivity. Took each frequency of each sensor as an unit, using the paired comparison method, calculated DI value of each unit distinguished by PCA (Principal Component Analysis, PCA). The ratio of the units with DI>0and the total number18which is confirmed as the distinction ability (Da%) of E-Tongue. When Da%has reached to50%or above, recorded the corresponding concentration minus the control as the difference threshold of E-Tongue sensors array.10different kinds of taste substances were detected by E-Tongue and got the threshold of each substance. It was compared with the human, the results as followed:sugar, caffeine and glycine detected by E-Tongue were less sensitive than human; quinine sulfate was similar with human; others such as citric acid, tartaric acid, acesulfame, sodium chloride, potassium chloride and sodium glutamate were more sensitive than human.
2. The concept of differential degree (Dd) is defined; it is tested by the distance between two classes. The higher the Dd value was, the greater difference existed, and vice versa. There are two differential degree test methods. The first one is the paired comparison test. There is a control sample R, which is stable properties belonging to the same category with the samples to be tested, and a series of the sample to be tested (1,2,3,4...... i), compared the test sample and the control sample respectively. The second one is sequential detection. Compared the relative Dd value between two samples respectively, or compared the samples and the control sample respectively.
3. Eight different kinds of basic taste substances were observed and a series of solutions were detected by E-Tongue with sequence order. The results showed that the Dd value was logarithmically increased with concentration, and the stimulus intensity increased at a geometric rate, whereas the differential degree increased at an arithmetic rate. This phenomenon was similar to Fechner's law in the field of psychophysics. The logarithmic relationship between the Dd value and concentration laid a theoretical foundation for E-Tongue detection.
4. Different brands of green tea beverage, and Chinese rice wine with different brands, different types and different ages were detected by E-Tongue and sensory evaluation. The results of the two ways were consistent.
The sensory attributes of green tea beverage such as tea flavor, sour, sweet, bitter were confirmed. Also the sensory attributes of Chinese rice wine such as ethanol, sour, sweet, bitter, astringent, pungent were confirmed. It can be evaluated the differential degree between two samples using Dd value.
The different brands of green tea can be identified by using DFA method with the correct rate of100%. The Shaoxing rice wine and non-Shaoxing rice wine can be identified by using SIMCA method with the correct rate of93.3%.
5. It was described a technique of shelf life analysis and quality control method based on E-Tongue test and sensory triangle test.
Chinese rice wine was detected by E-Tongue with sequence order, the Dd value of samples changed positively with exposure time. According to the result, it was considered that the Chinese rive wine was exposured for twelve hours as the quality control point, otherwise the quality was degrade. At the same time, it can be predicted the shelf life and quality based on the model of time and the Dd value. The orange juice beverage was detected by paired comparison test. The quality changed little at room temperature (24℃), but high temperature (37℃) can accelerate changes of the products, and the Dd value of samples changed positively with processing time. A quality control point was set for instance,5h, when Dd value was greater than2.67, the quality for orange juice beverage stored at37℃degraded.
6. The paper proposed the evaluation method of intelligent sensory instrument, such as electronic tongue and electronic nose, with the performance parameters as efficiency, repeatability, distinction stability, sensitivity and the detection limit. According to the evaluation method, the voltammetry E-Tongue was studied; at the same time, Smartnose and some brand electronic nose were studied. Smartnose and some brand electronic nose have there own advantages and disadvantages, need to be futhere improved.
本研究是基于智能感官领域相关研究存在的问题,着重从以下几个层面进行：(1)电子舌检测阈值和差别度理论研究,从仿生意义角度对经典心理物理学定律在电子舌物理传感器刺激响应方面的适用性进行研究,探讨基于差别度检验的电子舌检测理论依据；(2)规范出电子舌差别度检验的方法学框架,通过一定的手段将电子舌检测样品间的差别量化,建立样品间的差别度,科学的表征样品间的差别,并对电子舌检测不同样品的结果进行比较；(3)通过实例应用来对电子舌样品差别度表示方法的科学性进行验证；(4)对以电子舌和电子鼻为代表的智能感官仪器性能进行评价。本研究即从实际应用出发,解决电子舌应用中存在的实际问题。本课题主要研究结果如下：
1.定义了电子舌区分阈值的概念并规定了检测方法。差别阈为电子舌能够区分的样品的最小差别程度,即电子舌的灵敏度。将电子舌每个传感器每个频率段认为是一个单元,采用成对比较法,分别计算每个单元PCA区分的DI值,DI>0时是刚能区分开的临界值,而且是在高于该值时都能稳定的区分,所以认为当前的值是每个单元的差别阈值,再将各个单元综合起来,根据统计的每个单元的DI>0的数目与总的数目18的比值,为区分概率,即电子舌的区分能力Da%,当Da%达到50%或以上,所对应的浓度记为电子舌传感器阵列对该物质区分的差别阂值。根据上述方法分析和检测了10种味物质的差别阈值,并对电子舌与人的灵敏度进行了对比,通过比较可知,除了蔗糖、咖啡因和甘氨酸电子舌区分不如人灵敏外,对硫酸奎宁区分与人相近,其他的味物质如柠檬酸、酒石酸、AK糖、氯化钠、氯化钾、谷氨酸钠电子舌区分灵敏度要远高于人类。
2.定义了表征电子舌检测样品间差别度的概念、表示方法和差别度检验方法。
在几何空间上,对两个样品差别程程度大小的度量,即差别度(Differential degree, Dd),是通过计算两样品间的距离Da值来反映的,通过简单溶液分析,Dd值与物理刺激量呈现正相关,Dd值越大说明样品间的差别程度越大。Dd的引入将样品间不能度量的差别以具体数值的形式表现出来。建立了两种电子舌差别检验方法：一是成对比较检验法,确定一个对照样品R(性质稳定,且与所比较的样品属同一类)和一系列待检测样品(1,2,3,4……i),待检样品分别与对照样进行比较；二是顺序检测法,分别比较样品间的相对差别度,或将一系列样品分别与对照样进行比较。
3.通过对八种味物质溶液分析可知,电子舌检测样品的差别度Dd值与物理刺激强度间存在类似费希纳定律的现象。电子舌检测的样品Dd值与I(溶液浓度)间呈现对数关系,刺激强度按几何级数递增,而电子舌检测的样品间差别度按算术级数递增。电子舌检测样品间Dd值与I的关系与心理物理学研究中的费希纳定律类似,为今后电子舌的研究与应用奠定了一定的理论基础。
4.电子舌检测与感官品评技术结合实现了不同品牌绿茶饮料的区分,中国黄酒品牌区分、种类区分和不同年份区分,两种分析技术结果相互印证。
确定了茶味、酸度、甜度、苦涩感作为茶饮料感官品评的感官特性指标,确定醇香、酱香、酸、甜、苦、涩、辛辣感作为黄酒感官品评的感官特性指标。根据电子舌检测样品间的差别度Dd值大小可以判定样品间差别程度的大小。
对电子舌检测数数据采用DFA方法可以实现对不同品牌绿茶饮料进行辨识,辨识正确率100%；采用SIMCA方法可以实现绍兴黄酒与非绍兴黄酒的辨识,辨识正确率93.3%。
5.建立了产品货架期分析和质量控制与管理的电子舌差别检验和感官三点检验方法。
采用顺序检测法,分析了黄酒敞口放置时的质量变化情况。随着敞口放置时间的延长,Dd值有增大的趋势,即样品的差别度与时间具有正相关变化趋势。通过电子舌检测结合感官品评,12h可作为黄酒在室温下敝口放置的质量控制临界值,在12h或时间更长时,黄酒的感官质量明显变差,而且已经能够被人类所察觉。通过建立的Dd值与时间关系的数据模型可以对未知样品的质量和货架期进行分析预测,达到产品质量控制的日的。
采用成对比较法,分析橙汁饮料在不同条件处理时,根据Dd值判定其质量变化情况。窒温24℃处理对橙汁饮料的质量影响不大;而在高温37℃处理对其质量影响显著,Dd值随着处理时间的延长具有不断增大的趋势,将5h作为橙汁饮料在高温下质量控制临界点,当Dd值大于2.67时,产品就会发生较大的质量变化。
6.提出并构建了以检测效率、重复性、区分稳定性、灵敏性检测限这五个指标为标准的智能感官仪器(电子舌、电子鼻)性能评价方法,规范了操作方法和数据处理方法。依据构建的性能评价方法,对伏安电子舌的性能进行了研究；同时对本课题组开发的智鼻和某电子鼻的性能进行了研究和比较。
Electronic tongue (E-Tongue), one kind of intelligent sensory instruments, was produced with the development of sensory evaluation techniques, and in time, its applications have been increasing. This device is characterized by analysis of the overall characteristic, non-destructive process for the samples and designed for rapid detection. It has proven to be a good alternative for sensory evaluation techniques with objective and scientific, meets with the rapid development of the modern society. Less than30years since its development, it has been proven to be an ideal alternative for traditional chromatographic and in the analysis of food. Its great application potential has been expressed gradually; it has not only succeeded in distinguishing and identifying basic taste substances, but also in penetrating into various fields of food research. All of these studies have been mainly based on the principle component analysis (PCA) to distinguish and recognize substances qualitatively. Minimal research efforts in the quantitative expression of the sample difference have been undertaken. The purpose of the development and use of the E-Tongue is to make sensory evaluation scientific, objective, and quantitative, with the end view of using it in artificial sensory evaluation. Material distinction is not enough; the difference between samples must be quantified and resolved for a wider application of the E-Tongue in the future. Although researchers have proposed using a distance to characterize the relative difference between the two types of samples, and this research remains to be a concern worthy of further investigation.
This research was inspired by a related study in sensory field. In the intelligent sensory field, issues arise from the following aspects:first, the research of the threshold of the substances detected by E-Tongue, and the applicability of the classical law of psychophysics in E-Tongue and the exploration of the theoretical foundation based on difference test; second, the confirmation of the methodology used in the difference test for E-Tongue applications to qualify differences of the samples, establish the differential degree(Dd), characterize differences between samples scientifically, and compare the results of different samples; third, it is also the intent of the research to validate the science behind sample difference; forth, evaluate the performance of the intelligent instruments, such as E-Tongue and Electronic nose. This paper mainly embarked from the practical application angle, and resolved actual existing questions on the E-Tongue as a handy, rapid, and objective method. The results are showed as follows:
1. Established and confirmed the concept and test method of the threshold detected by E-Tongue. The difference threshold was defined as the smallest difference between samples distinguished by E-Tongue, and that was sensitivity. Took each frequency of each sensor as an unit, using the paired comparison method, calculated DI value of each unit distinguished by PCA (Principal Component Analysis, PCA). The ratio of the units with DI>0and the total number18which is confirmed as the distinction ability (Da%) of E-Tongue. When Da%has reached to50%or above, recorded the corresponding concentration minus the control as the difference threshold of E-Tongue sensors array.10different kinds of taste substances were detected by E-Tongue and got the threshold of each substance. It was compared with the human, the results as followed:sugar, caffeine and glycine detected by E-Tongue were less sensitive than human; quinine sulfate was similar with human; others such as citric acid, tartaric acid, acesulfame, sodium chloride, potassium chloride and sodium glutamate were more sensitive than human.
2. The concept of differential degree (Dd) is defined; it is tested by the distance between two classes. The higher the Dd value was, the greater difference existed, and vice versa. There are two differential degree test methods. The first one is the paired comparison test. There is a control sample R, which is stable properties belonging to the same category with the samples to be tested, and a series of the sample to be tested (1,2,3,4...... i), compared the test sample and the control sample respectively. The second one is sequential detection. Compared the relative Dd value between two samples respectively, or compared the samples and the control sample respectively.
3. Eight different kinds of basic taste substances were observed and a series of solutions were detected by E-Tongue with sequence order. The results showed that the Dd value was logarithmically increased with concentration, and the stimulus intensity increased at a geometric rate, whereas the differential degree increased at an arithmetic rate. This phenomenon was similar to Fechner's law in the field of psychophysics. The logarithmic relationship between the Dd value and concentration laid a theoretical foundation for E-Tongue detection.
4. Different brands of green tea beverage, and Chinese rice wine with different brands, different types and different ages were detected by E-Tongue and sensory evaluation. The results of the two ways were consistent.
The sensory attributes of green tea beverage such as tea flavor, sour, sweet, bitter were confirmed. Also the sensory attributes of Chinese rice wine such as ethanol, sour, sweet, bitter, astringent, pungent were confirmed. It can be evaluated the differential degree between two samples using Dd value.
The different brands of green tea can be identified by using DFA method with the correct rate of100%. The Shaoxing rice wine and non-Shaoxing rice wine can be identified by using SIMCA method with the correct rate of93.3%.
5. It was described a technique of shelf life analysis and quality control method based on E-Tongue test and sensory triangle test.
Chinese rice wine was detected by E-Tongue with sequence order, the Dd value of samples changed positively with exposure time. According to the result, it was considered that the Chinese rive wine was exposured for twelve hours as the quality control point, otherwise the quality was degrade. At the same time, it can be predicted the shelf life and quality based on the model of time and the Dd value. The orange juice beverage was detected by paired comparison test. The quality changed little at room temperature (24℃), but high temperature (37℃) can accelerate changes of the products, and the Dd value of samples changed positively with processing time. A quality control point was set for instance,5h, when Dd value was greater than2.67, the quality for orange juice beverage stored at37℃degraded.
6. The paper proposed the evaluation method of intelligent sensory instrument, such as electronic tongue and electronic nose, with the performance parameters as efficiency, repeatability, distinction stability, sensitivity and the detection limit. According to the evaluation method, the voltammetry E-Tongue was studied; at the same time, Smartnose and some brand electronic nose were studied. Smartnose and some brand electronic nose have there own advantages and disadvantages, need to be futhere improved.
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