| In this study the detection and removement technology of pyrogens as well as the freeze drying methods of polysaccharopeptide of coriolus versicolor were investigated.Firstly, We compare qualitative methods including common LAL gel-clot assay, special LAL gel-clot assay and LAL gel-clot assay with reagent of resist enhancing indicate that PSP interfere LAL test greatly, which result in the fake positive results. In the most valid diluted concentration range, all the experiments show positive results. Therefore, all these methods are not suitable for the detection of bacteria endotoxin because it has substance in PSP.The quantitative method including limulus test colorimetry which using the same system model method (using sample analog to replace the water without bacteria endotoxin as the reaction medium in limulus test experiment) to determine the bacteria endotoxin in PSP quantitatively. The result indicate the conelation coefficient is 0.997, which is over the value of 0.98; The calculation of recycle rate shows that the recycle rate is 104±10.9% at the concentration of sample diluted 300 times, according with criterion of the national pharmacopoeia for detection of endotoxin.All these results suggest that this limulus test colorimetry method is suitable for the detection of bacteria endotoxin in PSP. At the same time the experiment results suggest that the content of bacteria endotoxin is 2949EU/g in PSP. The value beyond the limit bacteria endotoxin of PSP (2400EU/g). Therefore, removing bacteria endotoxin in PSP is must.Secondly, methods used for removing pyrogen from PSP, including adsorption with active carbon, super-filtration and affinity chromatography as well as the effect of these methods on the physico-chemical characters including the content polysaccharide and protein, molecular weightdistributing and physiological activity of PSP including control cancer and immunocompetence in vitro was analyzed in this study. The result indicate that the content bacteria endotoxin of PSP reduce to 1773.9EU/g at the adsorption condition of pH 7.5, 90℃and 1% dosage in an hour. Although the adsorption method with active carbon to remove pyrogen from PSP can not affect the distribution of molecular weight of PSP, but affect the content of poly-glycoproteins, especially the content of protein, and reduce of the physiological activity in some different adsorption conditions.Using super-filtration method to remove pyrogen from PSP, obtain three substance, molecular weight beyond of 30kd, molecular weight under of 5kd and molecular weight in the middle of 30kd and 5kd. we find the substance of molecular weight under of 5kd of the content of bacteria endotoxin reduces to 322.6EU/g. But it seriously affect on the physico-chemical characters and physiological activity of PSP to use super-filtration method to remove pyrogen from PSP. Therefore it is not suitable for removing pyrogen from PSP, either.A good result was obtained when using affinity chromatography to remove pyrogen from PSP, the content of bacteria endotoxin in PSP reduce to 2301EU/g when the proportion sample: material is 5: 1 and recyled three times. Furthermore, it does not affect the content of poly-glycoproteins, the physiological activity and distribution of molecular weight of PSP. The result of statistic analysis shows that there no significant difference between the sample before and after affinity chromatography. Therefore it is the most suitable method to remove pyrogen from PSP.Furthermore, vacuum Freezing & Drying Technology was carried out in pilot study. To analyze from the shape of PSP have been freezed and dried, the best freezing and drying result was obtained with the load content of PSP at 1ml, under the concentration at 50mg/ml.and pre-freezed PSP with ethanol at minus 70℃and then pre-freezed with Ultra-low Temperature Freezer(-20℃) for 24 hours.Third provide a suitable technology and theory basis for research and develop the injection of PSP.
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