CN108489910B - 一种基于高光谱技术的牡蛎体内微塑料快速检测方法 - Google Patents
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Abstract
本发明属于食品安全检测技术领域，一种基于高光谱技术的牡蛎体内微塑料快速检测方法，步骤如下：(1)利用可见近红外光谱技术快速将微塑料从牡蛎组织以及其他体内杂质中区分开来；(2)利用光谱技术结合支持向量机等监督分类的方法可实现牡蛎体内不同类型微塑料的识别；(3)结合高光谱的图像技术和光谱技术，可以实现牡蛎体内微塑料空间分布的可视化；同传统的微塑料检测方法相比，基于高光谱技术的检测方法无需对生物体进行酸或碱等消解处理，可节省大量酸、碱试剂，减少环境压力。此外，基于高光谱技术的检测方法耗时短，并可实现微塑料空间分布的可视化。
Description
技术领域
本发明属于食品安全检测技术领域，涉及一种牡蛎体内微塑料快速检测的方法，具体是采用可见近红外高光谱技术对牡蛎体内不同的微塑料进行快速识别、检测，并实现牡蛎体内微塑料空间分布的可视化。
背景技术
近几年海洋环境中塑料垃圾堆积的问题引起越来越多人的关注。由于塑料自身的特性，难以降解的塑料在环境中会存在百年以上，因而会对环境造成持久的影响。海洋中的塑料在太阳辐射下产生光降解和破碎，形成粒径<5mm塑料，定义为微塑料。微塑料在环境中性质稳定，可存在百年甚至千年。海洋中的微塑料还可以富集多种有毒化学物质。海洋环境中的微塑料通过被海洋生物摄食，通过食物链的积累，甚至可以危害到人体的健康。因此，生物体内微塑料的检测是当务之急。目前海洋生物体内微塑料的检测主要包括两部分：(1)微塑料的提取；(2)微塑料物理形态和化学成分的检测。生物体内微塑料的提取是先利用酸性消解、碱性消解或酶消解等对样品进行预处理，从而排除生物组织的干扰。然后，利用饱和的食盐水使微塑料浮到溶液表面进行提取。微塑料物理形态的观察主要借助电子显微镜或扫描电镜等。而微塑料化学成分的确定主要借助红外光谱或拉曼光谱进行分析。
高光谱技术同时包含样本的光谱信息和空间图像信息。图像信息能对目标物的物理形态特征进行分析，光谱信息可以对目标物的化学成分进行鉴定。本发明开发了一种可见近红外高光谱技术对牡蛎体内的微塑料进行快速检测，并实现其体内微塑料空间分布可视化的方法。本发明拟结合化学计量学的方法，通过对高光谱图像上图像、光谱信号的分离来代替微塑料实际的分离提取过程，即采用“数理分离”代替“理化分离”，从高光谱图像上直接将微塑料从牡蛎体内识别，无需微塑料的提取过程，并实现微塑料在牡蛎体内空间分布的可视化。
发明内容
本发明的目的在于提供一种基于高光谱技术的牡蛎体内微塑料含量快速检测的方法。同现有的生物体内微塑料检测方法不同，高光谱技术可以实现牡蛎体内微塑料的快速检测，不需要对物体进行消解处理。且结合高光谱技术的图像特征，可以快速对微塑料的空间分布进行可视化。
本发明的技术方案：
一种基于高光谱技术的牡蛎体内微塑料快速检测方法，步骤如下：
将牡蛎内脏组织进行压片、风干处理，然后采集风干压片的可见近红外高光谱图像。采用支持向量机等监督分类方法，建立牡蛎内脏组织中微塑料的检测模型。最后，结合高光谱的图像特征，实现牡蛎内脏中微塑料分布的可视化。
所述的可见近红外高光谱的分析条件是指：光谱扫描范围为400-2500nm。
本发明的效果和益处：同传统的微塑料检测方法相比，基于高光谱技术的检测方法无需对生物体进行酸或碱等消解处理，可节省大量酸、碱试剂，减少环境压力。此外，基于高光谱技术的检测方法耗时短，并可实现微塑料空间分布的可视化。
附图说明
图1是高光谱采集系统图。
图2是微塑料的分类结果图。
具体实施方式
以下结合技术方案和附图，详细叙述本发明的具体实施方式。
一种基于高光谱技术的牡蛎体内微塑料快速检测方法，步骤如下：
(1)微塑料的准备
将几种常见的塑料材料：聚乙烯，聚丙烯，聚苯乙烯树脂，聚碳酸酯，聚对苯二甲酸乙二醇酯原料分别剪成粒径为0.1-5mm的微塑料若干，用不同目的筛子将微塑料按照尺度大小分成1-5mm、0.8-1mm、0.6-0.8mm、0.4-0.6mm、0.2-0.4mm以及0.1-0.2mm一共6个等级。
(2)牡蛎样本的准备
从市场上购买新鲜活的牡蛎样本，进行解剖，取其内脏、充分搅碎。取步骤(1)准备的每一粒径范围内每种微塑料各6个，将同一粒径范围内的5种塑料(总数30个)进行混合，共6个粒径尺度的混合微塑料样本。将同一尺度下的混合微塑料样品与搅碎后的内脏充分混合。
(3)含有微塑料内脏的前处理
为了便于微塑料高光谱信号的采集，本发现采用挤压的方法将步骤(2)中的内脏混合物挤压成薄薄的一片，厚度为0.1mm。由于水对可见近红外的光谱的干扰较大，本发明提出对挤压的样品在常温下进行自然风干处理。
(4)高光谱图像的采集
采集步骤(3)自然风干后样品的可见近红外高光谱图像，高光谱采集系统如附图1所示。采集光谱范围在400-2500nm。调整高光谱系统中摄像头距离样品的高度范围为11-13cm、曝光时间为1.4ms、帧数范围为641-758，在调试过程中选择最优的参数进行图像采集，获取最清晰高光谱图像；
(5)高光谱图像的处理
使用中值滤波(median filter,MF)、主成分分析(principal componentanalysis,PCA)、平滑(smooth)等方法对风干样品的高光谱图像进行平滑、去噪处理。从高光谱图像上选取各种不同类型微塑料以及牡蛎内脏组织的感兴趣区域，获取各种感兴趣区域的平均光谱曲线，对不同成分的光谱信号进行分析，采用支持向量机(Support vectormachine,SVM)等监督分类的方法对风干压片中的微塑料进行快速识别、分类。计算不同尺寸下，不同成分微塑料从内脏组织中的检出的概率。0.4-0.6mm样品检出率如附图2所示(高光谱图像上，每种塑料仅选取1粒作为相应感性区域的选取范围)。
采用传统检测方法对高光谱建立的方法进行验证
为验证该发明建立的方法是否可靠，风干的样本用酸性或碱性试剂消解12小时后，用饱和的食盐水提取消解液中的微塑料。采用拉曼光谱技术对提取出来的微塑料类似物进行成分鉴定。记录各风干样本中微塑料的数量，并将该结果与高光谱技术检测出来的微塑料数量进行对比，验证其准确性。
高光谱技术对实际样本中微塑料检测的应用
养殖含有微塑料的牡蛎，并用本发明的方法检测其体内微塑料的含量。养殖牡蛎用的容器为1000L钢化玻璃鱼缸，用海水洗刷后加满海水浸泡3-5天，然后用海水冲洗干净。将大约300L海水过滤，加入鱼缸中。从市场上购买活牡蛎若干。然后放入60只用海水洗刷干净的牡蛎。将微塑料与鱼饵混合，对活牡蛎进行喂食一周左右。每日换水，连续充气；一周之后，从实验池中取出牡蛎，利用步骤(1)-(5)建立的方法，对牡蛎内脏组织里的微塑料进行检测，并实现内脏组织中微塑料空间分布的可视化。
Claims (5)
1.一种基于高光谱技术的牡蛎体内微塑料快速检测方法，其特征在于，步骤如下：
(1)微塑料的准备
将塑料剪成粒径为0.1-5mm的微塑料，过不同目的筛子，得到不同尺度的微塑料；
(2)牡蛎样本的准备
将新鲜活的牡蛎进行解剖，取其内脏、充分搅碎；再加入步骤(1)得到的各尺度的微塑料搅碎，充分混合，形成多组含有微塑料的内脏混合物；
(3)含有微塑料内脏的前处理
将步骤(2)中的内脏混合物挤压成厚度为0.1mm的薄片，在常温下进行自然风干处理；
(4)高光谱图像的采集
采集步骤(3)自然风干后样品的可见近红外高光谱图像，采集光谱范围在400-2500nm；调整高光谱系统中摄像头距离样品的高度范围为11-13cm、曝光时间为1.4ms、帧数范围为641-758，在调试过程中选择最优的参数进行图像采集，获取最清晰高光谱图像；
(5)可见近红外高光谱图像的处理
对样品的可见近红外高光谱图像进行平滑、去噪处理；从可见近红外高光谱图像上选取各尺度的微塑料以及牡蛎内脏的感兴趣区域，获取感兴趣区域内所有像素点的平均光谱曲线，对光谱信号进行分析，采用支持向量机监督分类的方法对风干薄片中的微塑料进行快速识别、分类；
计算不同微塑料尺度下，不同成分微塑料从内脏组织中的检出的概率。
2.根据权利要求1所述的牡蛎体内微塑料快速检测方法，其特征在于，所述的塑料材料为聚乙烯、聚丙烯、聚苯乙烯树脂、聚碳酸酯或聚对苯二甲酸乙二醇酯。
3.根据权利要求1或2所述的牡蛎体内微塑料快速检测方法，其特征在于，所述的微塑料按照尺度大小分成1-5mm、0.8-1mm、0.6-0.8mm、0.4-0.6mm、0.2-0.4mm或0.1-0.2mm。
4.根据权利要求1或2所述的牡蛎体内微塑料快速检测方法，其特征在于，步骤(5)中，对于可见近红外高光谱图像进行平滑处理，采用的方法为中值滤波法、主成分分析法或平滑法。
5.根据权利要求3所述的牡蛎体内微塑料快速检测方法，其特征在于，步骤(5)中，对于可见近红外高光谱图像进行平滑处理，采用的方法为中值滤波法、主成分分析法或平滑法。
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| CN117554319A (zh) * | 2023-10-20 | 2024-02-13 | 广东省水利水电科学研究院 | 一种微塑料丰度的检测方法、系统、装置及存储介质 |
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