Studies On Chemical Composition's Isolation Analysis And Activity's Evaluation Of Compound Indigowoad Root Granule

The chemical components in traditional Chinese medicine (TCM) are responsible for their therapeutic effects and are very important in the modernization of TCM. High performance liquid chromatography (HPLC) and mass spectrometry (MS) techniques have been proved to be powerful tool in the analysis of TCM due to the high efficiency, sensitivity, speed, resolution and low level of sample consumption. Especially with the development of high performance liquid chromatography couple to mass spectrometry (HPLC-MS), it has provided direct, rapid and effective method for the authentication of known substances, structural identification of unknown substances and the selection of bioactive components in TCM.In this paper we separated different chemical composition by systemic solvent step by step extraction from compound Indigowoad Root granule and its single herbal medicine. Then we obtained the thirteen samples B1, B2, B3, B4, D1, D2, D3, D4, FF1, FF2, FF3, FF4 and FF.The analytical method of compound Indigowoad Root granule and its single herbal medicines using HPLC and MS had been preliminarily established. We also analyzed the chemical composition of all samples by HPLC-MS and other chemical identification techniques. So the mainly chemical components of compound Indigowoad Root granule and its single herbal medicine had been identified. The results showed that the mainly chemical components in the samples B1, D1 and FF1 are adenosine, uridine and alkaloids such as indigo, indirubin, hydroxy indirubin and tryptanthrin B. The mainly ones in the samples B2, D2 and FF2 are alkaloids such as 2,4(1H,3H)-quinazolinedione and 2,5-dihydroxy-indole and purine pyrimidine compounds such as guanine, adenine and uracil. The mainly ones in the samples B3, D3 and FF3 are epigoitrin and L-proline. And the mainly ones in the samples B4, D4 and FF4 are carbohydrates, amino acids and coumarin category.Then we studied on the biological activity of compound Indigowoad Root granule and its single herbal medicines by the evaluation in vitro. Through the impact of the compound Indigowoad Root granule and its single herbal medicines extracts on the proliferation of the isolated mouse spleen lymphocyte and the activity of the peritoneal macrophage, we found the effect of every extracts on the immune function of the isolated mouse. The results showed that every extracts can certainly stimulated the proliferation of spleen lymphocyte and activity of peritoneal macrophages. And the samples B3, B4, D4, FF3 and FF4 presented much stronger effect on the proliferation of spleen lymphocyte.The studies on the chemical composition and biological activity of compound Indigowoad Root granule and its single herbal medicine showed that the effective components of immunity enhancement are mainly existed in the larger polarity part, which are the carbohydrates, amino acids and coumarin category. The results were useful for the quality control of the compound Indigowoad Root granule and the bioactive material basis of this prescription.