Quality Standard Of Sambucus Sibirica Nakai And Application Of Infrared Spectroscopy In The Characterization Of Three Herbs

Objective:A quality control standard and method of Sambucus sibirica Nakai was developed by establishing several analysis methods including microscopic identification, thin-layer chromatography, high performance liquid chromatography. Using the near infrared spectroscopy and chemometrics method, three qualitative identification models were established for Fructus Cinnamomi cassiae immaturi, Peganum harmala L and Sambucus sibirica Nakai respectively which provided a new control standard and fast analysis method for identification of these medicinal material. Methods:1A microscopic identification method was developed for the study of Sambucus sibirica Nakai.10batches of Sambucus sibirica Nakai were analyzed the moisture, acid insoluble ash and total ash content. A TLC method was established for the qualitative identification of Sambucus sibirica Nakai using rutin and quercetin as internal control. The content of rutin, quercetin and betulinic acid in Sambucus sibirica Nakai from different batches were determined and compared by using HPLC method.2Using near infrared diffuse reflection spectroscopy combined with principal component analysis, clustering analysis, and SIMCA method, a qualitative model of Sambucus sibirica Nakai、Peganum harmala L、 Fructus Cinnamomi cassiae immaturi was established to fast identify these medicinal material. For fast quantitative analysis of the content of rutin in Sambucus sibirica Nakai, a near infrared diffuse reflection optical spectra method, together with partial least squares and principal component regression method, was established. Results:According to the analysis results of the established microscopic identification method, the recommended control standard of moisture, acid insoluble ash and total ash content of Sambucus sibirica Nakai is less than9.37%,0.71%and12.12%respectively. When rutin was used as the reference control, TLC identification method for Sambucus sibirica Nakai has high specificity in an ethyl acetate-formic acid-water (8:1:1) developing system. Meanwhile, when quercetin was used as the reference control, TLC identification method for Sambucus sibirica Nakai has high specificity in a toluene-ethyl acetate-formic acid (9:3:1) developing system. Using the established HPLC method, the content of rutin, quercetin and betulinic acid in Sambucus sibirica Nakai was determined. The content of rutin, quercetin and betulinic acid showed good linear relationship in200～1000μg-ml-1,0.8～8μgml-1,20-320μg·ml-1respectively (r=0.9999,0.9997,0.9999). The average recovery was100.38%,99.23%and99.61%respectively (RSD=2.04%,0.96%,1.34%, n=9).2According to the results of the full spectral analysis of the principle component analysis in Sambucus L.and Malaytea Scurfpea Fruit, the collected samples could be classified into four types. Using the principle component analysis method, Fructus Cinnamomi cassiae immaturi herb samples could be classified into six types and Peganum harmala L samples were classified into four types according to the origin of samples. A forecast model was established using clustering analysis method. The accuracy for Samucus sibirica Nakai, Fructus Cinnamomi cassiae immaturi and Peganum harmala L samples were95.24%,100%,91.67%respectively. A model established using SIMCA method was used for forecasting Sambucus sibirica Nakai, Fructus Cinnamomi cassiae immaturi and Peganum harmala L sample. The total recognition rate were85.71%,97.22%, and97.22%respectively. The analysis method of near infrared spectroscopy combined with partial least squares regression and principal component regression method was used for setting up Sambucus sibirica Nakai quantitative analysis model. The results showed that the root-mean-square errors of cross-validation of rutin were0.365and0.4476, the root-mean-square errors of calibration were0.2066and0.3791, and the predicted root-mean-square errors were0.2520and0.4291. The correlation coefficients of cross-validation were0.9792and0.9635. The correlation coefficients of calibration were0.9923and0.9746. The predicted correlation coefficients were0.9860and0.9596. All the results showed that our established method of the near-infrared spectroscopy could be used for the quantitative analysis of Sambucus sibirica Nakai. Conclusion:This study established a TLC identification method and a HPLC determination method of rutin, quercetin. and betulinic acid for the analysis of one of the Xinjiang local characteristics herb, Sambucus sibirica Nakai. The quality of Sambucus sibirica Nakai could be well evaluated by the established analysis methods. The near infrared spectroscopy together with chemometrics method was well applied to the qualitative analysis of Sambucus sibirica Nakai, Fructus Cinnamomi cassiae immaturi and Peganum harmala L which provided a new method for quality control of these herbs.In addition,these methods is also applied to the quantitative analysis of Sambucus sibirica Nakai.