目的: 利用表面增强拉曼光谱和化学计量学方法(surface enhanced laser Raman spectroscopy, SERS)对冬虫夏草(Cordyceps sinensis)和不同来源蛹虫草(Cordyceps militaris,柞蚕、大米培养蛹虫草和野生蛹虫草)进行定性研究,以建立一种可快速鉴定冬虫夏草和蛹虫草的方法。方法: 设置激光拉曼光谱仪的光谱测量范围0~3 200 cm-1,激光波长633 nm,总功率20 mW,物镜50倍,激发光源的强度为0.5%,扫描时间1 min,采集样品经表面增强技术处理前后的激光拉曼光谱,应用Origin 8.0软件对光谱进行降噪和归一化处理;进一步通过方差分析结合聚类分析和主成分分析,建立冬虫夏草与蛹虫草的品种判定方法。结果: 4种样品的SERS有7个共有峰:150、228、812、934、1 041、1 395、2 929 cm-1,其中冬虫夏草的150、228、812、934 cm-1 4个吸收峰强度显著弱于蛹虫草,而1 041、2 929 cm-1 2个吸收峰强度显著强于蛹虫草;3种蛹虫草在1 625~2 929cm-1范围内均有较强的吸收峰,而冬虫夏草则没有。主成分分析和聚类分析结果表明,3种蛹虫草归为一类,冬虫夏草为一类。结论: 本方法操作简便、快速准确、检测过程无损,所得图谱特征及区别明显,可用于冬虫夏草和蛹虫草的定性鉴定。
Objective: To establish a method for rapid identification of C.sinensis and C.militaris by surface-enhanced Raman spectroscopy(SERS) and chemometric methods to qualitatively study C.sinensis and C.militaris from different sources(tussah, rice-cultured C.militaris and natural C.militaris). Methods: The spectral measurement range of the laser Raman spectrometer was 0-3 200 cm-1, the laser wavelength was 633 nm, the total power was 20 mW, the objective lens was 50 times, the intensity of the excitation light source was 0.5%, and the scanning time was about 1 min. The laser Raman spectra of the samples before and after surface enhancement technology were collected and Origin 8.0 software was used to denoise and normalize the spectra. Furthermore, a method for judging the species of C.sinensis and C.militaris was established by means of variance analysis combined with cluster analysis and principal component analysis. Results: The SERS of the four samples had seven common peaks at 150, 228, 812, 934, 1 041, 1 395 and 2 929 cm-1. The absorption peaks of C.sinensis (150, 228, 812 and 934 cm-1) were significantly weaker than those of C.militaris, while the absorption peaks at 1 041 and 2 929 cm-1 were significantly stronger than that of C.militaris. All the three species of C.militaris had strong absorption peaks in the range of 1 625-2 929 cm-1, while C.sinensis didn’t have. The results of principal component analysis and clustering analysis showed that the three species of C.militaris were classified as one type and C.sinensis as one type. Conclusion: The method is easy to operate, fast and accurate, the detection process is non-destructive, the characteristics and differences of the obtained spectra are obvious, and it can be used for the qualitative identification of C.sinensis and C.militaris.
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