| The compositions of traditional Chinese medicines (TCMs) are complicated, and the information of those compositions could provide basis for their pharmacology, medicinal efficacy, quality control and metabolism. So the separation analysis of those compositions in TCMs is very important. Nucleosides as a part of the basic substances in TCMs demonstrated a unique and irreplaceable role in anti-virus and anti-tumor sides, which can be used as drugs directly or indirectly. So the researchers had paid much attention to it. In this paper, the separation and analysis of nucleotides in Dragonhead and Bulbus Fritillariae were investigated, and the results could provide direction for the further study.In addition, due to the difference of the property and structure of different compositions in different TCMs, the separation of different compositions needs to investigate the effect of various factors of HPLC condition, such as the composition of mobile phases, types of stationary phase and column temperature. Effect of the selection of mobile phase for separation of TCMs was investigated in this paper. We chose Salvia, Atractylodes and Coptis Chinensis which have different acid-base properties about their composition as the example drugs and the peak capacity, peak profile and resolution were used as the evaluation indexes.The details are summarized as follows:1. To develop HPLC-UV-ESI/MS method to detect the nucleosides and bases of Dragonhead. In the paper, we chose Dracocephalum Tanguticum Maxim as the raw material to optimize the extracting and separation analysis conditions. Under the optimum conditions, qualitative and quantitative analysis to the nucleosides of the Dracocephalum Tanguticum Maxim and Dracocephalum argunense were performed. The method is accurate, quick and highly effective, which can be used as the basis of the quality analysis of Dracocephalum.2. A high-performance liquid chromatography method was developed for simultaneous determination of five nucleotides in Bulbus Fritillary. Meanwhile, the quality analysis and the species difference of the nucleosides from the different sources were discussed. The method is simple, convenient and accurate. It can be used for the determination of nucleosides and supplying evidence for exploiting and applying of Bulbus Fritillariae.3. In the paper, reversed-phase high performance liquid chromatography (RP-HPLC) was used for the separation of TCMs in gradient mode. We screened different kinds of mobile phase according to solvent selectivity triangle and investigated the influence of different kinds of mobile phase on the separation of the acidic, neutral and alkaline compounds in TCMs. Three typical TCMs(Salvia, Atravtylodes and Rhizoma Coptidis) were selected as analysis models. All of them were extracted by the basic solvent extraction method. The experiment was performed on a Spherigel ODS C18 column (200x4.6 mm,5μm) and in order to achieve the optimal separation of different kinds of component, we compared different mobile phase systems, modifiers, and changed its ionic strength and pH. The results not only provided the chromatographic separation conditions for these kinds of Chinese medicinal materials and the study on fingerprint chromatography about them, but also provided the basis for the classification of the unknown compounds in TCMs.
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