| ObjectiveUnder the instruction of theory of Chinese Medicine, we carried out a systematic study on Microctis Folium of Genuine Regional Chinese Materia Medica in Guangdong Province. The modern analytical techniques were used to establish the genuine regional quality Evaluation and clarify the Lipid-regulating pharmacodynamics components. We expect the study will lay a scientific basis for its resources development and clinical application, as well as provide a new research idea and method for studying Genuine Regional Chinese Materia Medica in Guangdong Province.MethodsMicroctis Folium from different regions were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) fingerprint, the first derivative ultraviolet (UV) spectroscopy and Fourier transform infrared (FTIR) spectroscopy, in order to establish the chromatogram and spectrum fingerprint. Total flavonoids were determined by UV Spectrophotometry for rutin control, while the contents of vitexin and isovitexin determined by HPLC. Multivariate calibration model between the UV spectroscopy and the extracts was developed by partial least squares (PLS), which is a new method for rapid determination of the extracts in Microctis Folium. By observing the total cholesterol (TC) and triglyeeride(TG) in serum, Lipid-regulating effect of total flavonoids, alkaloids, tannins and total organic acids in Microctis Folium were evaluated by mice of hyperlipidemia, which induced by Triton WR-1339.ResultsResults of fingerprint:Observing TLC fingerprint, Microctis Folium from different regions have the same chemical composition, while the contents are different. TLC of kaempferol has clear spots and good separation, by using silica gel G plate prepared1%sodium hydroxide solution and developing solvent of toluene-ethyl acetate-formic acid (9:3:1), while TLC of flavonoids of vitexin, isovitexin and narcissoside using silica gel G plate and ethyl acetate-methanol-water (100:17:13) solvent system, which coming out good effect. The HPLC fingerprint study results show that the different regions Microctis Foliumhave high similarity in HPLC fingerprint. The identification of different regions Microctis Folium can be enabled by similarity evaluation software. The cluster analysis method of UV derivative spectrum can identify Microctis Folium and the other four leafy Chinese medicines accurately, and reflect the relationship between each sample. It provides a fast, accurate and feasible identification method for Microctis Folium Seventeen characteristic peaks were found on IR fingerprint. Comparing the overall shape and characteristics of absorption peaks in spectrum, it can be achieved the purpose of rapid identification.Result of study on determination of Microctis Folium from different regions:Microctis Folium from different regions has different contents of total flavonoids, vitexin and isovitexin, while total flavonoids is between2.61%to14.86%, vitexin is0.028%to0.289%, and isovitexin is0.060%to0.469%. The changes of vitexin and isovitexin are in the same direction, which is different from total flavonoids. Results suggest that the quality of Microctis Folium was influenced by environment, processing and storage conditions, and so on. It should use the multi-index comprehensive evaluation for quality control. The result of this study can provide an experimental basis for optimal selection of materials.The study of UV-PLS for the determination of extracts in Microctis Folium:Model calibration coefficient (R2) is0.9641, while root mean square error (RMSE) is0.6931. The predicted results show that the method can predict the content of extracts accurately, with the average relative deviation of1.73%, respectively. It shows good predictive ability, and provides a reference for rapid quality control in pharmaceutical process.Pharmacodynamic study:Compared with negative group, model group shows a significant increase in TC and TG (P<0.01). The groups of total flavonoids and alkaloids have a significant decrease in TC and TG (P<0.05or P<0.01), when compared with the model group. The results suggest that total flavonoids and total alkaloids in Microctis Folium have a significant role in regulating the experimental hyperlipidemia in mice, while total flavonoids was more significant (P<0.01).Conclusions:On the basis of existing quality standards of the "Chinese Pharmacopoeia", this subject promoted its level of quality control. It established the database of TLC and HPLC fingerprint, UV and FTIR spectrum, and the determination methods of total flavonoids, vitexin and isovitexin for Microctis Folium. It also proposes a new method (UV-PLS) for a rapid determination of extracts in Microctis Folium. The pharmacological experiments indicated that total flavonoids and total alkaloids have a significant role in regulating the experimental hyperlipidemia in mice, which laid a good foundation for health care and treatment. |
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