| Honeysuckle is one of common clinic traditional herb, the present studies show Chlorogenic acid is the most effective component, which has marked activities such as detoxification, diminish inflammation, antisepsis and so on. The thesis had studied the extraction, isolate and purification process of Chlorogenic acid by comparative methods, on top of, investigated the biological activity of extractions and identified the composition in the extractions. Results showed:1.Extraction process of Chlorogenic acid8 fold of 70% grain alcohol, refluxing 2 hours, in twice, reflux conditions: pH 4 or so, under 60 ; The total extractions rate is less than 33%, extraction rate of Chlorogenic acid is above 92%, and the content of Chlorogenic acid is about 18.58%.2. Isolate & purification process of Chlorogenic acidThe process is combined with two steps: first, primary enriched and isolated chlorogenic acid from the extractions of the honeysuckle by precipitation with Zn2+; second, advanced isolated and purification chlorogenic acid from the former more pure substance through the technology of macroporous adsorption resin (JAD-1400), and thesame time, eliminating the abio-ions brought by the first step.Zn2+ precipitation conditions: optimum pH is 5.6, in the extraction solution which has 200mg chlorogenic acid per ml, amount volume of 1M ZnCl2 is complied with the rule of equation: Y=7.78314+43.39277 X (Y is the precipitation rate of chlorogenic acid, X is the amount volume of 1M ZnCl2); dissolving conditions: pH of the acid liquor is 1.5, the volume of acid liquor is 3 fold as much as the volume of chlorogenic acid precipitations, 60 dissolving 15 min, the received rate of chlorogenic acid is about 81.35%, the content of chlorogenic acid is over 40%.The conditions of purification with JAD-1400 Resin:Resin sorption: confecting saturated solution of extractions(600ml), preparing pH 3, and adding NaNO3 till 7% of concentration, inflooding saturated solution into column resin in 12ml/min(resin column:3.5cm 80cm, the volume of resin : 721.3cm3); Resin isolation: first, 1200ml of distilled water inpoured the column in apt velocity of flow; second, 900ml of 20% ethanol flowed into the column at 15ml/min; following, the afflux of 900ml 45% ethanol with grads velocity, gathering the deep ingredient; finally, regenerating the resin with 95% ethanol and water; at last, we gained more pure canary matter, content of chlorogenic acid is above 83%, the gained rate of chlorogenic acid inexcessof 38%.3. Identifying the composition in the extractionsUtilizing the measure of Thin Layer Chromatography(TLC) assay, High Performance Liquid Chromatography(HPLC) assay , Ultraviolet- Visible light Scanography(UV-S) assay to identify the composition in the extractions. From the fingerprint chromatogram of assay, we can draw a conclusion that chlorogenic acid is the most composition in the extractions. Indicating our process stable and safe.4. Investigating the biological activity of extractionsResearch showed: Chlorogenic acid has antioxidation, eliminating free radicel and antisepsis action. In the antioxidation, high pure chlorogenic acid is better than low pure one, but they are weaker than tea plyphenols; in the eliminating free radicel, chlorogenic acid has select eliminating ability, significant difference with different content, IC50 (DPPH, OH)of high pure CHA is 65mg/l and 135 mg/1, and low one is 327mg/l and 1432mg/l, but tea plyphenols is 52mg/l andl 908 mg/1; in the antisepsis: chlorogenic acid can inhibit both staphylococcus aureus and escherichia coli. .The MIC of high pure chlorogenic acid is twice as the one of low pure chlorogenic acid at staphylococcus aureus; at escherichia coli., they have no difference. Tea plyphenols is more effect than chlorogenic acid in the antisepsis.
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