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The 916 is a marine marine sulfated polysaccharide chitin as raw materials, multi-step carboxymethylation and sulfation the molecular modification from polyanion sulfate glycosaminoglycan deacetylation. System pharmacodynamics, pharmacology, toxicology studies, found that the 916 has a good anti-atherosclerosis activity, has been approved by the State Drug Administration as a new class of drugs I enter clinical studies. The thesis aims for 916 human pharmacokinetic study provides preliminary of work the investigated 916 under certain experimental conditions, the stability and establish a HPLC method for in biological samples rabbit serum. Select two enzymes present in biological fluids in the stability studies, --- lysozyme and trypsin as the conditions of the inspection, the use of high performance gel permeation chromatography (HPGPC), and gelatin turbidimetric method, were investigated different indicators, from a molecular weight and molecular skeleton is changed to the active group is off in order to determine the stability of 916. 916 molecular weight and molecular weight distribution coefficient visit HPGPC law, the effective group sulfuric acid ester group stability investigated using gelatin free sulfate content than the turbidity quantitative detection samples. The results show that is stable under the selected experimental conditions, the 916 pairs of lysozyme and trypsin may be presumed, in the serum 916 will not be degraded by these two enzymes. To further study the stability of the body 916 in a previous treatment of the biological sample and the biological mechanisms, certain preparations. Determination of derivatization rabbit serum concentration of 916 papers for the first time the use of HPLC columns, select the guanidine hydrochloride as derivative reagent, and to explore the optimization of the derivative reaction conditions is. Comprehensive comparison of several analytical column as NH2 column Asahipak NH 2 P-50 4E gel column TSK-Gel. G2000PW, the effect of the use of the Aminex HPX-87H ion exclusion column, established ion exclusion column Aminex HPX-87H analytical column of sulfated polysaccharide 916. In the biological sample of rabbit serum pretreatment, the application of a variety of inter the protein method such as a protein precipitating agent, trichloroacetic acid, perchloric acid, zinc salts, acetonitrile, methanol, ethanol, a saturated ammonium sulfate, etc. protein precipitation method, an ultrafiltration method and trypsin protease digestion and compared acetonitrile precipitation method for sample processing method, the establishment of a standard curve and linear range, determined 916 in the rabbit serum, the detection limit and quantitation limit, 2.5μ g / ml and 10μg/ml respectively. studied precision, reproducibility, and recovery of rabbit serum 916 HPLC method provides a methodological basis for 916 preclinical and clinical pharmacokinetic studies.
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