Optimization of Ultrafiltration Technology of Enzymatic Hydrolysate from Eucommia ulmoides Peel / 中国药房

OBJECTIVE：To optim ize the ultrafiltration technology of enzymatic hydrolysate from Eucommia ulmoides peel. METHODS：The single factor test was adopted to investigate the effects of molecular weight of ultrafiltration membrane ，liquid temperature，operating pressure ，operating frequency ，membrane filtration time ，liquid concentration and pH on transfer rates of aucubin，geniposide and chlorogenic acid as well as solid removal rate in enzymatic hydrolysate from E. ulmoides peel. Setting the molecular cut off of fixed ultrafiltration membrane of 100 000，liquid concentration of 7 g/L，and pH value of 7，the ultrafiltration technology was optimized by Box-Behnken design response-surface methodology and validated with liquid temperature ，operating pressure，operating frequency and membrane passing time as factors ，using comprehensive scores calculated from transfer rates of aucubin，geniposide and chlorogenic acid as well as solid removal rate as indexes. RESULTS ：The optimal ultrafiltration technology of enzymatic hydrolysate from E. ulmoides peel was as follows as liquid temperature of 35 ℃，operating pressure of 0.5 MPa，operating frequency of 35 Hz and membrane passing time of 42 min. Results of validation tests showed that the comprehensive scores of the transfer rates of aucubin ，geniposide and chlorogenic acid as well as solid removal rate in enzymatic hydrolysate from E. ulmoides peel was 78.06％（RSD＝1.43％，n＝3），and its relative error with the predicted value （77.18％） was 1.14％. CONCLUSIONS ：The optimized ultrafiltration technology is stable and reliable ，and can be used for the ultrafiltration purification of enzymatic hydrolysate from E. ulmoides peel.

Structure and application of Ganoderma polysaccharide peptide GL-PPSQ2 / 中草药

Objective To study the structure of active polysaccharide peptide purified from Ganoderma lucidum aqueous extract, and used as a reference substance for the determination of polysaccharide peptide content in G. lucidum products. Methods GL-PPSQ2 was obtained by hot water extraction, separation and purification with membrane ultrafiltration and gel-filtration chromatography. The physicochemical determination and spectral date were used for structural identification. The content of polysaccharide peptide was detected by HPLC with UV detector, water was used as mobile phase and the flow rate was 1 mL/min. Results GL-PPSQ2 was a pure polysaccharide peptide with purity above 97%, molecular weight of 5.0 × 104, polysaccharide content of 87.17%, and yield of 0.49%. The monosaccharides composition analysis showed that GL-PPSQ2 was glucose-based polysaccharide with a small amount of mannose, which contained 16 kinds of amino acids with the total amount of amino acids of 5.04%. Based on the methylation analysis, 1D and 2D NMR spectroscopy, the repeating unit of GL-PPSQ2 was composed of →3)-β-D-Glcp-(1→backbone, with four repeating units connected a long chain branch at O-6 which was composed of α-D-Glcp-(1→, →4,6)-β-D-Glcp-(1→, →4)- β-D-Glcp-(1→ and →6)-β-D-Glcp-(1→ in sequence. Conclusion The active polysaccharide peptide was isolated and purified by membrane technology and gel chromatography. The method was simple and rapid, which provided a scientific basis for the quality control of polysaccharide peptide in G. lucidum extract and its products.

Membrane technology for optimizing process optimization of small bioactive compounds in Mailuoning Injection factory effluene / 中草药

Objective: To enrich small bioactive compounds from Mailuoning Injection factory effluene by polyether sulfoxide hollow fiber membrane. Methods: The retention rate of the common polymer (protein, tannin, starch, and pectin) and the permeability of the small molecule pharmacodynamic substance chlorogenic acid were the indicators, and the stop time and membrane type in ultrafiltration process were optimized. The temperature, pressure, and feed rate were investigated to determine the influence degree of each factor on the permeability and retention rate, and then the optimum process parameters of the ultrafiltration membrane process were optimized by using Box-Behnken design-response surface method. Results: The sequence of common polymer retention rate from high to low is: the influencing factor A (temperature), B (pressure), C (feed rate), and there was a negative correlation with the temperature. For the chlorogenic acid transmission rate, the influencing factor A (temperature) was larger than B (pressure), and C (feed rate) was lowest. There was a “∩” relationship between chlorogenic acid permeability and pressure at a certain temperature and rate. The results of optimized process were as follows: liquid temperature was 35 ℃, feed rate was 262 mL/min, and membrane pressure was 15.0 psi (1 psi = 6.895 kPa). The chlorogenic acid transmission rate was (91.470 ± 0.988)%, and polymer retention rate was (80.080 ± 2.296)%. The comprehensive score was (85.780 ± 1.641)% and RSD was 0.46%. There was a small difference between the two processes. Conclusion: Hollow fiber membrane, which is a sustainable and green resource utilization, has a good permeability for small bioactive compounds in wastewater, and has excellent retaining effect on common macromolecules.

Optimization of ultrafiltration technology for water extract in Cornus officinalis / 中草药

Objective: To explore the possibility for the application of ultrafiltration technology using polysulfone (PS) hollow fiber membrane to purlfy the water-extraction of Corni Fructus and optimize the preparation procedure. Methods: The PS membrane of different aperture 5×104, 1×104, and 6×103 were studied on the applicability with water-extraction of Corni Fructus. The proper molecular weight retention was optimized with the removal of liquid impurities and the transformation. Taking the ratios of loganin transmittance and common polymers (pectin, tannin, protein, and starch) retention as indexes, the proper ultrafiltration condition was optimized by response surface method. Results: The PS membrane with interception molecular weight of 1×104 had better separation efficiency and the preferred operation parameters were 0.1 MPa of pressure, 50℃ of liquid temperature, and solution concentration of 0.13 g/mL. Conclusion: The preferred aperture of PS membrane and the operation conditions are feasible for the purification of water-extraction of Corni Fructus, which can improve the production efficiency.

Evaluation on effect of ultrafiltration technology on endotoxin removal from recombinant MUC1-MBP fusion protein / 吉林大学学报(医学版)

Objective To evaluate the effectiveness of ultrafiltration technology in endotoxin removal from purified recombinant MUC1-MBP fusion protein (MUC1-MBP)and to demonstrate the effect of ultrafiltration on endotoxin removal.Methods CM Sepharose FF weak cation exchange (CM)(CM group), CM combined with Phenyl Sepharose 6 FF exchange (C6)(CM+C6 group),CM combined with ultrafiltration (CM+ultrafiltration group), and CM combined with C6 and ultrafiltration (CM+C6+ultrafiltration group)were used to purify the MUC1-MBP from E.coli. and remove endotoxin;the expression level of endotoxin was detected by Chromogenic End-point Tachypleus Amebocyte Lysate.Results There was a single band at the expected molecular weight of 62 000 by SDS-PAGE analysis.and the purity>96% by Quantity One analysis.The endotoxin levels in CM group and CM +C6 group were quite high and there was no significant difference between two groups (P>0.05 );the endotoxin level in CM+ultrafiltration group was significantly lower than that in CM group, and there was significant difference (P0.05 ). Conclusion The effects of CM or CM combined with C6 on endotoxin removal are quite poor, especially C6;CM combined with ultrafiltration are quite effective on endotoxin removal,and ultrafiltration plays an important role in endotoxin removal.

Purification technology for decarburization fluid of Reduning Injection by polysulfone ultrafiltration membrane / 中草药

Objective: To investigate the ultrafiltration effect of polysulfone hollow fiber ultrafiltration membrane and polysulfone plate ultrafiltration membrane with relative molecular weight by different cut-off on the decarburization fluid of Reduning Injection. Methods: The transfer rates of chlorogenic acid and geniposide, fingerprint similarity, and removal of bacterial endotoxin, protein, oxalate, and resin before and after ultrafiltration were used as inspect indicators to optimize the purification technology for the decarburization fluid of Reduning Injection. Results: The obvious effect of polysulfone hollow fiber ultrafiltration membrane with the relative molecular weight 10000 on oxalate and resin in the decarburization fluid of Reduning Injection was observed. Bacterial endotoxin (100%) in the decarburization fluid of Reduning Injection was removed by the polysulfone plate ultrafiltration membrane with the relative molecular weight 5000. Conclusion: The stability and the preparation safety of Reduning Injection could be ensured by the two stage combination of the ultrafiltration membrane technology.

Optimization of ultrafiltration technology for Xiangdan Injection / 中草药

Objective: To explore the effect of ultrafiltration technology in the preparation of Xiangdan Injection on transmittance and impurity removal rates of protocatechuic aldehyde (PA) and sodium danshensu (SD), to evaluate its applicability, and to optimize the ultrafiltration technology. Methods: Taking the solution temperature, pH value, intercept relative molecular weight of ultrafiltration membrane, membrane import and export pressure difference of ultrafiltration membrane as influence factors, the process parameters of the ultrafiltration were optimized by detecting the contents and solid contents of PA and SD before and after the ultrafiltration. Results: The three factors of temperature, pH value, and import and export pressure difference have no significant influence on the transmittance of PA and SD, and the removal rate of impurity. The intercept relative molecular weight of ultrafiltration membrane has the significant influence (P < 0.01). Conclusion: The optimized ultrafiltration conditions are feasible for the preparation of Xiangdan Injection.