基于智能手机识别的薄层色谱法快速检测油菜籽中的叶绿素  

Rapid detection of chlorophyll in rapeseed by thin layer chromatography based on smartphone recognition

作　　者：郭梦帅 李泽一 肖华明 吕昕[1] 梅德圣[1] 王丹 姚旋 郭亮 胡琼[1] 陈洪[1] 魏芳[1,4] GUO Meng-shuai;LI Ze-yi;XIAO Hua-ming;LYU Xin;MEI De-sheng;WUANG Dan;YAO Xuan;GUO Liang;HU Qiong;CHEN Hong;WEI Fang(Oil Crops Research Institute of Chinese Academy of Agricultural Sciences,Key Laboratory of Oilseeds Processing,Ministry of Agriculture and Rural Affairs,Hubei Key Laboratory of Lipid Chemistry and Nutrition,Wuhan 430062,China;National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070,China;Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China;Hubei Hongshan Laboratory,Wuhan 430062,China)

机构地区：[1]中国农业科学院油料作物研究所,农业农村部油料加工重点实验室,湖北省油料脂质化学与营养重点实验室,湖北武汉430062 [2]华中农业大学,作物遗传改良国家重点实验室,湖北武汉430070 [3]农业农村部油料作物生物学与遗传育种重点实验室,湖北武汉430062 [4]湖北洪山实验室,湖北武汉430070

出　　处：《中国油料作物学报》2025年第1期217-225,共9页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(U21A20274);国家重点研发计划专项(2021YFD1600103);农业农村部油料作物生物学与遗传育种重点实验室开放课题(KF2023008);中国农业科学院创新工程项目(CAAS-ASTIP-2013-OCRI);湖北省自然科学基金创新群体项目(2023AFA042);农业农村部中央级公益性科研院所基本科研业务费(161072023021)。

摘　　要：为高效鉴定油菜籽品质,建立一种操作简单、成本低、耗时短的油菜籽中叶绿素含量的快速检测方法。采用乙醇(95%)提取样品中的叶绿素,以石油醚(60~90℃)-丙酮-甲苯(体积比2∶1.5∶2)为展开剂,紫外线分析仪(365 nm)激发叶绿素荧光显色,用智能手机采集荧光斑点图像,Image J软件进行荧光斑点自动识别,得到斑点面积,实现叶绿素a和叶绿素b的快速定性定量分析。该方法定量检测叶绿素a和b的线性范围为0.04~1.00 mg/m L,叶绿素a的决定系数R^(2)为0.9965,日内精密度为5.9%,日间精密度为8.6%。叶绿素b的决定系数R^(2)为0.9925,日内精密度为6.1%,日间精密度为5.6%。油菜籽中叶绿素a和叶绿素b的加标回收率分别为90.00%~91.67%和92.00%~113.00%。结果表明该方法线性关系较好,准确度良好,能快速测定油菜籽中叶绿素含量。For high efficient rapeseed quality detection,a method was developed on low-cost and time-efficient detecting chlorophyll content in rapeseed.In this method,extraction of chlorophyll from seed samples was achieved using ethanol(95%),followed by development with petroleum ether(60-90℃)-acetone-toluene(2:1.5:2,volume ratio).Chlorophyll fluorescence color was excited using a UV analyzer(365 nm).Fluorescent spot image was captured using a smartphone.Image J software was employed for automatic identification of fluorescent spots,measurement of spot area,and realization of rapid qualitative and quantitative analysis of chlorophyll a and chlorophyll b.The linear range for chlorophyll a and b was determined to be 0.04-1.00 mg/mL,with R^(2) of 0.9965 for chlorophyll a and 0.9925 for chlorophyll b.The intra-day precision was 5.9%for chlorophyll a and 6.1%for chlorophyll b,while the inter-day precision was 8.6%for chlorophyll a and 5.6%for chlorophyll b.The recoveries of chlorophyll a and b in rapeseed were observed to be between 90.00%and 91.67%for chlorophyll a,and between 92.00%and 113.00%for chlorophyll b.These results indicated the well linearity and accuracy of this method,enabling rapid determination of chlorophyll content in rapeseed.

关 键 词：叶绿素 薄层色谱法 Image J 智能手机 定量分析 

分 类 号：S565.4[农业科学—作物学]