Study on Antioxidant Components of Tibetan Medicine Sanguo Tang San
|School||Chengdu University of Traditional Chinese Medicine|
|Keywords||Tibetan medicine Sanguo Tang San Antioxidant Active components TLC-bioautography Structure identification|
Tibetan medicine Sanguo Tang San as the basic preparation in Tibetan medicine, consisted with Terminalia chebula Retz., T.billerica (Gaertn.) Roxb. and Phyllanthus emblica L. Fructus Chebulae, which can clear the blood-heat and dissolve the bad blood. Sanguo Tang San, is used for the treatment of plateau diseases. Modern studies have found that plateau disease are caused by metabolic imbalances of free radicals imposed by plateau hypoxia and series of pathological changes whereby from oxidative stress induces apoptosis. Therefor, the research below has revealed the antioxidant active components of Tibetan medicine Sanguo Tang San:Objective:Find a method to preliminarily clarify the antioxidative ingredients of Sanguo Tang San, which is mainly based on TLC-Bioautography, DPPH and HPLC.Method:1. To microscopic identify Sanguo Tang San and each single herb.2. To determine values of free radical-scavenging capacity, presented as DPPH, of Sanguo Tang San and each single herb by spectrophotometry.3. To screen the antioxidative ingredients in Sanguo Tang San by TLC-Bioautography techniques.4. To separate the ethyl acetate extracts of Fructus Phyllanthi by silica gel column chromatography, gel-column chromatography, preparative TLC, etc., and identify with modern spectrum technologies of 1H-NMR and 13C-NMR. And then to determine the values of radical-scavenging capacity of each separated ingredients.5. To determine gallic acid, the stronger antioxidative ingredient, as the index component in Sanguo Tang San by HPLC.6. To preliminarily analysis the constituents absorbed in blood by the HPLC fingerprints in vitro and vivo of Sanguo Tang San.Results: 1. The microscopic characteristics, including calcium oxalate crystal, non-glandular hairy, oil droplet,ect., supplemented the monograph of Sanguo Tang San in Chinese Departmental Pharmaceutical Standard·Tibetan medicine (1995’s).2. The discrepancies of Sanguo Tang San from different sources were little; and each single herb had strong antioxidative ability.3. Gallic acid, as the main composition in Sanguo Tang San, and ethyl acetate extracts in Frucrus Phyllanthi were showed strong antioxidant activity by TLC-Bioautography.4. Eight components of Fructus Phyllanthi (β-sitosterol,β-daucosterol, cinnamic acid, gallic acid, ethyl gallate, ellagic acid, mucic acid dimethyl ester-2-O-gallate and corilagin) were isolated and identified, among which the gallic acid had the stronger antioxidant activity.5. The determination of gallic acid in Sanguo Tang San were performed by HPLC., and the linear range was 0.1605～0.6420μg (r=0.9999).6. The serum fingerprints showed that gallic acid and corilagin were the prototype constituents absorbed in blood, and it revealed that they might be the direct antioxidant materials in vivo.Conclusion:Gallic acid, ethyl gallate, ellagic acid and corilagin have strong antioxidant ability in vitro. Combined with the serum fingerprints, gallic acid and corilagin have been preliminarily comfirmed as the antioxidative components of Sanguo Tang San.