首页|基于表面增强拉曼光谱的牛肝菌主成分分析及快速鉴定

基于表面增强拉曼光谱的牛肝菌主成分分析及快速鉴定

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2021年《中国疾病预防控制中心周报》指出蘑菇中毒是我国最严重的食品安全问题之一,2021年发生蘑菇中毒事件327次,涉及923例患者,其中20例死亡,总死亡率为2.17%,事件中共鉴定出毒蘑菇74种.牛肝菌由于味道鲜美,是人们最喜爱的菌类之一.由于烹调不当或是混入有毒牛肝菌,就会发生牛肝菌中毒事件.如何快速得出牛肝菌的主成分并进行鉴定,成为了一个迫切需要解决的问题.该研究从楚雄市鸿福农贸市场上购买的10个新鲜牛肝菌样品,使用菌盖进行徒手切片,用微波法制备的纳米银溶胶制成银胶装片,置于DXR显微拉曼光谱仪的10倍物镜下,获得10个样品的表面增强拉曼光谱(SERS).通过对SERS的分析发现10个样品中,样品7、8、10主成分相同,样品6、9主成分相同,其余样品主成分各不相同.以样品1为例,主成分有L-苯基丙氨酸(1 583 cm-1环内C—C伸缩振动、1 199 cm-1 NH2摇摆振动)、L-组氨酸(1 572 cm-1 C=C伸缩振动、COO-不对称伸缩、1 229 cm-1面内环变形振动)、异亮氨酸(1 342 cm-1C—H、N—H变形振动、486 cm-1 COOH摇摆振动、353 cm-1骨架振动)、L-天冬氨酸(1 136 cm-1 C—N伸缩振动)、甘氨酸(1 386 cm-1 COO-变形振动、889 cm-1 C—C伸缩振动)、蛋氨酸(681 cm-1 C—S反对称伸缩振动)、吡喃糖化幌物(973 cm-1对称环振动).采用光谱软件OMNIC Specta的建库功能随机挑选10个样品的一条谱线进库,建库后可快速进行光谱匹配度的检索,通过匹配度的高低鉴定待测样品.在10个样品中,除2、10号样品的匹配度小于60%外,其余样品的匹配度都大于60%.样品7、8的匹配度接近且有交叉,说明样品7、8是同一品种,而样品6、9的匹配度接近而交叉项少,说明这两个样品是同一品种或相近品种.DNA检测结果表明样品7、8、10同为白牛肝菌,而样品6、9同为薄瓤假美柄牛肝菌.该实验为牛肝菌的主要成分检测和快速鉴定提供了一种简单可靠的方法.同时,该方法对野生蘑菇的物种鉴定具有很大的应用潜力,在野生菌中毒病例中,迅速确定毒蘑菇种类,从而为抢救病人赢得时间具有重要的意义.为此首次对野生蘑菇子实体进行SERS研究.
The Main Ingredients Analysis and Rapid Identification of Boletuses Based on Surface-Enhanced Raman Spectroscopy
According to a 2021 China CDC Weekly Express report,mushroom poisoning was one of China's most serious food safety issues.In 2021,a total of 327 mushroom poisoning incidents involving 923 patients and 20 deaths were investigated,and the overall mortality was 2.17%.About 74 poisonous mushrooms have been successfully identified.Boletuses are one of the people's favourite wild mushrooms because of their delicious taste.However,boletuses poisoning incidents can occur due to improper cooking or mixing with toxic boletuses.Quickly identifying variety and the main ingredients of boletus has become a problem that needs to be resolved immediately.Thus,tenfresh boletuses samples were purchased from the Chuxiong Hongfu market.The pileus was prepared by free-hand section,then the silver colloid area was prepared by using the silver colloid prepared by microwave.Under the DXR Laser confocal micro Raman spectrometer,the surface-enhanced Raman spectroscopy(SERS)spectra of 10 samples were achieved.In these results,the main ingredients of samples 7,8 and 10 were the same,while samples 6 and 9 also shared the same main ingredients.However,the other samples had different ingredients compared to each other.As an example,in sample 1,the main ingredients were L-phenylalanine(1 583 cm-1 ring C—C stretching vibration and 1 199 cm1 NH2 rocking),L-histidine(1 572 cm-1 C=C stretching vibration COO-asymmetric stretching and 1 229 cm-1 in-plane ring deformation vibration),isoleucine(1 342 cm-1 C—H,N—H deformation vibration,486 cm-1 COOH rocking vibration and 353 cm-1 lattice vibration),L-aspartic acid(1 136 cm-1 C—N stretching vibration),glycine(1 386 cm-1 COO-deformation vibration and 889 cm-1 C—C stretching vibration),methionine(681 cm-1 C—S antisymmetric stretching vibration)and pyranose(973 cm-1 symmetric ring vibration)By applying the spectrum software OMNIC Specta and randomly selecting 10 lines to build the database of the sample spectra,the species of fresh boletuses could be identified quickly by measuring their spectra and matching them with the standard spectra in the database.In all samples,only samples 2 and 10 had relatively low matching rates,which were less than 60%.Furthermore,samples7 and 8 shared similar matching rates and cross terms with each other,which indicated they were of the same species.Samples 6 and 9 had similar matching rates but fewer cross terms,which indicated they were of the same or similar species.DNA test results showed that samples 7,8 and 10 were Boletus baingan,while samples 6 and 9 were Baorangia pseudocalopus.This experiment provides a simple and reliable method to detect the analysis of the main ingredients and rapidly identify Boletuses.Also,this approach has great potential for species identification of wild mushrooms.This experiment has great application value in quickly determining the poisonous wild mushroom species and gaining time to rescue the patient in the case of wild mushroom poisoning.To our knowledge,it is the first time SERS has been used on wild mushroom fruiting bodies.

BoletusNano-silverSurface-enhanced Raman spectroscopyMain ingredientMatching rate

司民真、王敏、李伦、杨永安、钟加菊、余成敏

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楚雄师范学院云南省高校分子光谱重点实验室,云南楚雄 675000

楚雄师范学院光谱应用技术研究所,云南楚雄 675000

楚雄彝族自治州人民医院蘑菇中毒临床研究室,云南楚雄 675000

牛肝菌 纳米银 表面增强拉曼光谱 主成分 匹配度

国家自然科学基金云南省科技计划科技重大专项

113640012018ZF009

2024

光谱学与光谱分析
中国光学学会

光谱学与光谱分析

CSTPCD北大核心
影响因子:0.897
ISSN:1000-0593
年,卷(期):2024.44(6)
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