ABSTRACT
Objective To optimize the water extraction and alcohol precipitation process of Radix Paeoniae Rubra-Cortex Moutan drug pair(1∶1).Methods L9(34)orthogonal experiment was used.The ratio of material to liquid,extraction time and times of extraction were taken as the factors,and the extraction amount of paeoniflorin and paeonol and the yield of dry extract were taken as the indexes.Hassan's method was used for multi-index comprehensive scoring to optimize the water extraction process.The extract density,ethanol volume fraction and standing time were taken as the factors,and the transfer rate of paeoniflorin and paeonol and the yield of dry extract were taken as the indexes.At the same time,the alcohol precipitation process was optimized according to the actual situation of large-scale production.Results The optimal extraction process was to add 10 times of water and decoct for 1 time,60 min for each extraction.The optimal alcohol precipitation process was as follows:the density of the liquid was 1.20 g·mL-1(30℃),95% ethanol was added to make the alcohol content up to 60% ,and the liquid was left standing for 60 h.Conclusion The optimized process of water extraction and alcohol precipitation is stable,reliable and with good repeatability,which can provide reference for the follow-up study of Radix Paeoniae Rubra-Cortex Moutan drug pair.
ABSTRACT
<p><b>OBJECTIVE</b>To obtain high yield and good antigenic activity of HDV L-Ag and to detect different regional patients' sera to test the purified antigen's antigenicity.</p><p><b>METHODS</b>Hepatitis delta virus' sequence was obtained from Inner Mongolian patient by using RT-PCR and PCR methods, PET43a was used and His-tag was added at the HDV L-Ag 5' and 3' to construct the recombinant expression plasmid, transform the plasmid into host bacterium BL21 and induce it with IPTG. The expression supernatant was purified by saturated (NH4)2SO4 and affinity chromatography. The activity and antigenicity of the expressed product were analyzed by using EIA.</p><p><b>RESULTS</b>Comparison of results obtained with detection by using the expressed protein coated plate and ABBOTT Murex anti-Delta (total) of 15 positive and 10 negative sera, the consistency was good (100%).</p><p><b>CONCLUSION</b>EIA proved that the purified antigen had good antigenicity, no serological difference was found in detection between different region's sera, therefore the purified delta antigen may be useful in diagnostic and other research.</p>