Study On The Preparing Technique Of The Effective Part And Its Tablet Of Polygonum Cuspidatum

AIMSOur aims were (1) to establish the determination method of the effective ingredients in Polygonum cuspidatum; (2) to optimize the extraction procedure for effective ingredients in Polygonum cuspidatum; (3) to study the technological parameters of the purification process for effective part from Polygonum cuspidatum; (4) to optimize the preparing technique of concentrated tablet of Polygonum cuspidatum (CTPC); (5) to analyze the effects of CTPC on liver injury of mice induced by CCl4; (6) to study the protective effect of CTPC on cyclophosphamide injured mice.METHODS1. The contents of polydatin, resveratrol, emodin, physcion and total anthraquinone were determined by HPLC and differential spectrophotometric method, respectively. With the extraction rates of the above ingredients as the parameters, the extraction conditions of Polygonum cuspidatum were optimized by orthogonal design.2. Using adsorption capacities and desorption rates of polydatin, resveratrol, emodin, physcion and total anthraquinone as the primary screening indexes, six resins were surveyed, and the optimized conditions of adsorption and desorption of the effective ingredients were studied.3. Using the disassembly time limit, absorption and facies of the tablets as the primary screening indexes, the formulation of CTPC was optimized, and the contents of the effective ingredients in the tablets were determined by HPLC and differential spectrophotometric methods.4. After administrating CTPC (2.5 g/kg, 1.25 g/kg, 0.25 g/kg) over 7 days consecutively, the mice were given CCl4 to lead to acute liver injury. The activities of serum ALT and AST, as well as the pathological changes of the liver were determined in all groups.5. Cyclophosphamide was ip into mice pretreated with CTPC and the level of leucocyte were observed.RESULTS1. The extracting temperature and time possessed notable effects on the comprehensive extraction of the active ingredients in Polygonum cuspidatum. The optimal procedure was obtained as follows: the sample was refluxed three times at 90℃with 5 times of 70% EtOH, 2 h for each time.2. Resin D101 gave good separation performance and was selected to purify the effective part in Polygonum cuspidatum. The optimum parameters were established as the following: 1 BV (bed volume) sample extract was passed through the column with a flow rate of 2.4 BV/h, 30 min later, the column was washed with 2 BV water, 2 BV 20% ethanol, 5 BV 50% ethanol, 2 BV 70% ethanol and 5 BV 95% ethanol, respectively. The combined 50% and 95% ethanolic elutes were concentrated to yield the purified effctive part.3. The optimum formulation of CTPC was composed of major medicine (total effective part of Polygonum cuspidatum) 0.195 g, lactose 0.08 g, MCC 0.10 g, CMC-Na 0.02 g and magnesium stearate 0.002 g. The tablets prepared was much better in hardness and facies. The determination results of the contents accorded with the regulation.4. CTPC could inhibit the rise of serum ALT(P<0.01) and AST(P<0.01) in mice with liver injury induced by CCl4, and relieve the degeneration and necrosis of liver tissues.5. CTPC obviously inhibited the decrease of leucocyte on cyclophosphamide injured mice.CONCLUSIONThe extraction and purification process of the effective part from Polygonum cuspidatum, together with the preparing technique of its tablets (CTPC) were studied systematically, with the major effective ingredients all adopted as evaluation indexes for the first time. The optimal extraction pocedure is rational, economical, with high yield of effective ingredients. Macroporous resin D101 could be well used in separating and purifying the effective part, with the high purity of the total effective ingredients in the product (36.87%). The tablet of the effective part prepared with the optimum formulation accorded with the regulation. The phamacodynamic studies suggested CTPC has protective effects on the acute liver injury induced by CCl4 and inhibit the decrease of leucocyte on cyclophosphamide injured mice. All the results suggested that the preparation of CTPC be successful and meaningful to developing a new preparation of Polygonum cuspidatum.