| A kind of analytical method by high performance liquid chromatography (HPLC) for tea catechins was established. In the meantime, the method was investigated in the methodological aspect through determining the main catechins EGCG of tea polyphenols. The optimum chromatographic conditions were as follows: ODS column (5μm,4.6×250 mm),distilled water (0.05% orthophosphoric)/ acetonitrile/ ethyl acetate is 86: 12: 2 (V/V) as mobile phase (pH=3.0), UV wavelength is 280 nm, the flow rate is 0.8 ml/min. The analytical method is simple, precise and easy to operate.The ester catechins was enriched containing EGCG, GCG, ECG from the raw TP in the pretreatment process and employed as raw stuff in purification process. Single factor experiment method was used to systematically study the separation and purification conditions of EGCG by liquid-liquid partition chromatography. Took silica gel as inert stuffing with the height of 40 cm and ether as elution agent with the elution speed of 5 ml/min in the separation process, and collect an elution every 4 minutes, we effectively obtained EGCG with high-purity. Besides, this method needed low cost, produced EGCG with high purity and achieved gram magnitude for once time.Tea polyphenols with the EGCG content about 60% were used as the raw materials, used the liquid-liquid partition column chromatography and choosed silica gel as the inert supporter to amplify the result uniformly based on the experiment condition, which was carried out in a small scale. As a result, we got a good method of separation and purification to obtain EGCG from the partition column: the deionized water was used as stationary phase, and ether was used as mobile phase, the elution speed was 10 ml/min, the ratio diameter to height was 1:10, collect the eluted liquid every 30 minutes. In addition, based on the experimental data, the stability of ester catechins was researched to provide scientific proof for storage of the finished product. In this work, more economic equipments had been used and EGCG of high purity can be obtained by using single partition column chromatography. The developed technology was simple and easy to control with the advantages of low cost. The separation technique was easy to be industrialized.The injection formulation and lyophilizing process was studied in the pharmic research on EGCG. The formulation and lyophilizing process are optimized by the criterion of appearance, water content and resolubility. The study showed that the formulation was 25 mg EGCG, 300 mg lactose and the specification was 2 ml each bottle. The optimized technological process was as followed: the drug was pre-freezed for 5 h under -40℃and dried for 32 h in the lyophilizer under–25.0℃. After that, the temperature was raised to 0℃to continue drying for 5 hours. The quality evaluation of final product was researched including the general examination items (clarity, pH, water content), the content and the stability experiment. Three months tests under room temperature or under accelerated circumstance indicate that the preparation was stable. |
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