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目的建立含量测定方法,对拉曼光谱内标法中2种参量的定量分析结果进行比较,确定拉曼内标定量法最适合的参量。方法分别以拉曼峰的相对峰强和峰面积比值作为参量,以不同采收时期和不同产地的连翘叶为对象,采用内标法进行连翘苷的含量测定。以相对峰强为参量时,选取样品溶液拉曼光谱中连翘苷苯环振动(1 319.1 cm-1处的强峰)和溶剂甲醇中CH3-O反对称伸缩振动(2 974.7 cm-1)作为定量峰和内标参比峰;以峰面积比值作为参量时,选取连翘苷苯环振动(1 319.1 cm-1处的强峰)和溶剂甲醇中CH3-O对称伸缩振动(3 020.9 cm-1)作为定量峰和内标参比峰。分别以定量峰与内标峰强和峰面积的比值为纵坐标,以连翘苷浓度为横坐标绘制标准曲线,并对线性和加样回收率进行比较。结果以2种参量作为响应值用于拉曼光谱定量分析,均有良好的线性,相关系数r分别为0.998 0和0.997 6;回收率均在100.04%～101.30%之间,回收完全;对2种参量的测定结果进行线性拟合,得出回归方程C2=1.030 4C1-0.033 1,r=0.999 6。结果表明两者测得结果一致,利用2种参量进行定量分析都能取得准确结果。结论无论以相对峰强,还是以峰面积比值作为参量都可以用于拉曼光谱定量分析,2种参量的测定结果基本一致。拉曼光谱用于连翘苷的定量分析具有操作简便、迅速等优点,可以用于中草药的含量测定。
OBJECTIVE To establish a method for the determination of content and compare the results of quantitative analysis of two parameters in Raman spectroscopy internal standard method to determine the most suitable parameters for Raman internal standard quantification. Methods The relative peak intensities and peak area ratio of Raman peak were used as parameters respectively. Forsythia suspensa leaves from different producing periods were collected for determination of forsythin by internal standard method. When the relative peak intensity is taken as the parameter, the vibration of the benzene ring of forsythin (strong peak at 1 319.1 cm-1) and the anti-symmetric stretching vibration of CH3-O (2 974.7 cm-1) (Peak at 1 319.1 cm-1) and CH3-O symmetric stretching vibration (3 020.9 cm-1) in methanol solvent were used as the parameters of peak area ratio and internal standard reference peak. -1) as the quantitative peak and internal reference peak. The ratio of quantitative peak to internal standard peak intensity and peak area was taken as ordinate, and forsythin concentration was taken as abscissa to draw a standard curve. Linearity and sample recovery were compared. The results of two kinds of parameters as the response value for the quantitative analysis of Raman spectroscopy, have a good linearity, the correlation coefficient r were 0.998 0 and 0.997 6; recoveries were between 100.04% ~ 101.30% The results of the species parameters were linearly fitted and the regression equation C2 = 1.030 4C1-0.033 1, r = 0.999 6 was obtained. The results show that the two measured results are consistent, the use of two kinds of parameters for quantitative analysis can get accurate results. Conclusion Both the relative peak intensity and the peak area ratio can be used for the quantitative analysis of Raman spectra. The results of two kinds of parameters are basically the same. Raman spectroscopy for forsythin quantitative analysis has the advantages of simple, rapid, etc., can be used for the determination of Chinese herbal medicine.