1.Determination of total content of ten ginsenosides in yiqifumai lyophilized injection by near infrared spectroscopy.
Xiaoping HAN ; Dekun LI ; Dazheng ZHOU ; Ruichao LIN ; Lifang LIU ; Zhengliang YE
China Journal of Chinese Materia Medica 2011;36(12):1603-1605
OBJECTIVETo determine the total content of 10 ginsenosides in Yiqifumai lyophilized injection by near infrared spectroscopy.
METHODSixty samples were collected and determined of the total contents of ten ginsenosides by HPLC. The optimal calibration model was established by the contents of 10 ginsenosides in fifty samples and their NIR spectroscopy using the PLS. And the contents of 10 samples were successfully predicted.
RESULTWhen using the pretreatment of the first derivative and MSC in the range of 4 246.8 - 4 602.2, 5 446.8 - 61 02.6 cm(-1), the best dimension was 9, and the quantitative model was accurate. The R2 was 94.2, and the RMSECV was 0.186. The RMSEP of ten samples was 0.234.
CONCLUSIONThis method is easy, rapaid and precise, and can be used to determine the content of 10 ginsenosides in Yiqifumai lyophilized injection.
Freeze Drying ; Ginsenosides ; chemistry ; Spectroscopy, Near-Infrared ; methods
2.Development of a freezing drier for lyophilization of biomaterials.
Zhigu WU ; Miao GEN ; Tongzhu SUN ; Xiaobing FU ; Dexin XIONG ; Yingwu QIAO ; Yuejin SONG ; Shixue SUN ; Zhiyong SHENG
Journal of Biomedical Engineering 2004;21(3):460-463
To observe and assess the performance and effect of our self-made FD-1 freezing drier on biomaterials. R502 compressor and R502 refrigerating agent were adopted. In the experiment, FD-1 lyophilized collagen sponge, strain and defibrinogenase. The evaporating-condenser temperature reached -45 degrees C and the small icebox temperature reached -30 degrees C under the loading or free-loading circumstances in the lyophilizing box. The lyophilized collagen sponge had many pores in the structure, and the strain and the defibrinogenase were lyophilized and maintained satisfactorily. This freezing drier is suitable for lyophilizing some biomaterial samples in small or medium batches.
Bacteria
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Collagen
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Freeze Drying
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instrumentation
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methods
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Humans
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Temperature
3.Optimization of lyophilization procedures for freeze-drying of human red blood cells.
Lin-feng CHEN ; Jing-han LIU ; De-qing WANG ; Xi-lin OUYANG ; Yuan ZHUANG ; Ji CHE ; Yang YU ; Hui LI
Journal of Southern Medical University 2010;30(9):2055-2058
OBJECTIVETo investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells (RBCs).
METHODSHuman RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 degrees celsius;. The morphological changes of the RBCs were observed to assess the degree of vitrification, and the specimens were placed in the freeze-dryer with the temperature set up at 40, -50, -60, -70 and -80 degrees celsius;. The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared, and the water residue in the specimens was determined.
RESULTSThe protectant did not show ice crystal in the course of freezing and thawing. No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of -70 degrees celsius; and -80 degrees celsius; (P > 0.05). The E procedure resulted in the maximum recovery of the RBCs (83.14% ± 9.55%) and Hb (85.33% ± 11.42%), showing significant differences from the other groups(P < 0.01 or 0.05). The recovery of the RBCs showed a positive correlation to the water residue in the samples.
CONCLUSIONFast cooling in liquid nitrogen and shelf precooling at -70 degrees celsius; with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.
Erythrocytes ; cytology ; Freeze Drying ; Humans ; Tissue Preservation ; methods
4.Aggregation after rehydration of lyophilized platelets.
Jing-Han LIU ; Fa-Qiang LU ; Yuan ZHUANG ; Ji CHE ; Lin-Feng CHEN
Journal of Experimental Hematology 2006;14(4):812-815
This study was aimed to investigate the aggregation of rehydrated-lyophilized platelets. The aggregation rate of fresh and rehydrated-lyophilized platelets were measured by using thrombin, ristocetin, ADP and collagen as inductors and APACT2 aggregameter; the effects of intra- and extra-cellular trehalose on maximum aggregation rate of rehydrated-lyophilized platelets were detected by using ADP as an inductor. The results showed that the aggregation rate of fresh platelets was all about 100%, while aggregation rate of rehydrated lyophilized platelets was (70.17 +/- 7.36)%, (15.3 +/- 2.81)%, (68.67 +/- 6.86)%, (64.67 +/- 11.6)% respectively, when the concentration of thrombin, ristocetin, ADP and collagen was 1 U/ml, 1.6 mg/ml, 20 micromol/L and 2 microg/ml. The maximum aggregation rates of rehydrated-lyophilized platelets in intra- and extra-cellular trehalose, extracellular trehalose and blank control groups were (66.0 +/- 4.69)%, (25.3 +/- 2.42)% and (11.5 +/- 1.87)% (P < 0.01), meanwhile there was significant difference of rehydrated-lyophilized platelet aggregation rate between intra- and extra-cellular trehalose and extracellular trehalose groups (P < 0.01). It is concluded that the concentrations of thrombin (1 U/ml), ristocetin (1.6 mg/ml), ADP (20 micromol/L) and collagen (2 microg/ml) are optimal for platelets aggregation tests, the internal and extracellular trehalose significantly enhance the aggregation of rehydrated-lyophilized platelets.
Blood Platelets
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cytology
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drug effects
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metabolism
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Blood Preservation
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methods
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Freeze Drying
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methods
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Humans
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Platelet Aggregation
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physiology
5.Effects of different lyophilizing protectants on lyophilized trehalose-loading red blood cells.
Yan CHEN ; Zhi-Gang LU ; Hai BAI
Journal of Experimental Hematology 2013;21(3):775-779
This study was purposed to evaluate the effect of different lyophilizing protectants including human albumin, glucan, polyvinyl pyrrolidone and glycerine on lyophilized trehalose-loading red blood cells (RBC), then to screen the optimal lyophilizing protectant. The RBC were incubated in 800 mmol/L concentration of trehalose solution at 37°C for 7 hours, and washed 3 times with PBS solution to obtain the trehalose-loading RBC. The trehalose-loading RBC in control group were directly lyophilized without lyophilizing protectants, the trehalose-loading RBC in the experimental group were mixed with Lyophilizing protectants. The samples of 2 groups were kept at room temperature for 30 minutes, pre-frozen at -80°C for 24 hours, then lyophilized in freeze-dryer for 24 hours. Finally the samples were quickly rehydrated by 6% HES at 37°C. The recovery rate and hemolysis rate of hemoglobin were detected by using cyanohemoglobin detection kit. The water content of unhydrated samples were detected at the same time. The results showed that when the moisture content of sample was 3% - 5%, the recovery rate of hemoglobin in control group was 33.57 ± 2.89%, and that in experimental group was 51.15 ± 1.98%, there was statistically significant difference between the control and experimental group (P < 0.05). When the different concentration of dextran solution was chosen as protectants, the recovery rate of hemoglobin of lyophilized RBC was obviously lower. The higher concentration of dextran, the better the recovery rate. The recovery rate of hemoglobin was 22.15 ± 4.12% when the concentration of dextran was 36%, there were statistically significant difference between the two groups (P < 0.05). When the different concentration of polyvinyl pyrrolidone (PVP) solutions was chosen as protectants, especially the concentration below 40%, the recovery rate of hemoglobin of lyophilized RBC was significantly belower than the control group, there was statistically significant difference between the two groups (P < 0.05). When 10% glycerol was used as protectants, the recovery rate of hemoglobin was 3.93 ± 1.80%. There was also statistically significant difference between the two groups (P < 0.05). It is concluded that human serum albumin shows an important protective effect on the lyophilization of the trehalose-loading red blood cells. The dextran and PVP at the concentration lower than 40% can decrease the protective effect of trehalose in cells. Glycerol can not be chosen as protectant for lyophilized trehalose-loading red blood cells.
Blood Preservation
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methods
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Cryoprotective Agents
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pharmacology
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Erythrocytes
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drug effects
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Freeze Drying
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methods
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Humans
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Trehalose
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pharmacology
6.Optimization for vacuum belt drying process of Panax notoginseng extract.
Xue-Song LIU ; Zhi-Fang QIU ; Long-Hu WANG ; Yi JI ; Yi-Yu CHENG ; Hai-Bin QU
China Journal of Chinese Materia Medica 2008;33(4):385-388
OBJECTIVETo optimize the conditions of the vacuum belt drying process (VBD) for drying Panax notoginseng extract and compare with methods of vacuum freezing drying and spray drying.
METHODThe optimum conditions of VBD were obtained by orthogonal design and validated by determinations of moisture content of the dried product and recovery of active ingredients. Experiments on different drying methods were also conducted.
RESULT AND CONCLUSIONThe optimum conditions are as follows, the feeding speed was 15 mL x min(-1), the belt speed was 4 mm x min(-1), and the heating temperature was (105, 100 degrees C). Comparing with the drying methods of vacuum freezing drying and spray drying, vacuum belt drying possesses some advantages, such as higher recovery of active ingredients, less moisture content of dried product and better overall yield.
Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Freeze Drying ; instrumentation ; methods ; Vacuum
7.Preliminary study on rehydrated conditions for lyophilized human red blood cells.
Lin-Feng CHEN ; Jing-Han LIU ; De-Qing WANG ; Xi-Lin OUYANG ; Yuan ZHUANG ; Ji CHE ; Hui LI
Journal of Experimental Hematology 2009;17(6):1582-1587
The objective of this study was to investigate the effect of different rehydration conditions on recovery of the lyophilized red blood cells (RBC) so as to optimize the RBC rehydration. The different conditions, including different rehydration solution, the rehydration temperature, volume change rate of the lyophilized RBC rehydrated by the vapor firstly, were studied, the recovery rate and change of physiological and biochemical properties of the rehydrated RBC were detected. The results indicated that the solution of 10% (w/v) PVP40 in PBS showed the best effect, and the RBC recovery rate increased with increasing of rehydration temperature, and the optimal temperature of rehydration was at 37 degrees C. Pre-rehydration in condition of vapor could raise the RBC recovery rate, and promote the MCV and RDW to close to index of the fresh RBC, the deformability of the rehydrated RBC was no serious as compared with RBC preserved in conventional condition, but the activity level of ATP, G-6-PD, SOD, 2, 3-DPG of the rehydrated RBC less decreased. It is concluded that the optimal rehydration conditions for lyophilized RBC are pre-rehydration in the 37 degrees C with vapor firstly, PBS + 10% (w/v) PVP40 rehydration solution and rehydration temperature at 37 degrees C, but the protection of RBC membrane needs to be furtherly studied.
Blood Preservation
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methods
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Erythrocyte Count
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Erythrocytes
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Freeze Drying
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methods
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Humans
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Rehydration Solutions
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Temperature
8.Effects of freeze-drying process on polymerized human placenta hemoglobin.
Lanzhen ZHAO ; Shen LI ; Fengjuan LI ; Jinfeng WANG ; Chengmin YANG
Journal of Biomedical Engineering 2013;30(5):1052-1057
The present study was aimed to investigate the influence of freeze-drying on the quality of polymerized human placenta hemoglobin (PolyPHb). The PolyPHb solution was freeze-drying under suitable conditions. Hemoglobin concentration, methemoglobin (MetHb) content, UV spectrum, Fe3 content, oxygen-carrying capacity, pH, the average molecular weight and its distribution, circular dichroism, oxygen equilibrium curve and other indicators were measured before and after freeze-drying. The appearance, residual water content, rehydration time of the lyophilized product were also evaluated. The results showed that there was no significant difference on all the indicators measured above, which indicated that freeze-drying process had no effect on the physical and chemical properties of PolyPHb, as well as on its biological activity. Therefore, the properties of PolyPHb were stable during this freeze-drying process and could be preserved after such freeze-drying process.
Blood Substitutes
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Female
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Freeze Drying
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methods
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Hemoglobins
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chemistry
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Humans
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Methemoglobin
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analysis
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Placenta
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chemistry
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Polymerization
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Pregnancy
9.Trehalose -- a biomembrane protectant applied to lyophilization of human red blood cells: review.
Journal of Experimental Hematology 2006;14(5):1061-1064
Trehalose, as a nonreducing disaccharide, plays a role in protecting the cytoactivity when the cells is freezing, drying or lyophilization. It has been a biomembrane protectant applied to lyophilization of human blood cells (platelets and erythrocytes), and from which astonishing results have been obtained. Having powerful hydration, distinctive vitrification transform and crystal transform and unique resistance of high temperature and humidification, trehalose is thought of a preferred protectant in the study of cell preservation. In recent years, people concerned trehalose on its protective mechanism, experimental means of transit trehalose to mammal cells and the mechanism of loading in red blood cells. The above aspects were briefly summarized in this article.
Blood Preservation
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methods
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Cryoprotective Agents
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pharmacology
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Erythrocytes
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cytology
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drug effects
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Freeze Drying
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Humans
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Trehalose
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pharmacology
10.Effect of trehalose on the freeze-dried boar spermatozoa.
Xiangqian MENG ; Xiaolong GU ; Caifeng WU ; Jianjun DAI ; Tingyu ZHANG ; Yini XIE ; Zhiqiang WU ; Liang LIU ; Hengdong MA ; Defu ZHANG
Chinese Journal of Biotechnology 2010;26(8):1143-1149
After freeze-drying, the ultrastructure of boar sperms was observed by optical and electron microscopy. The in vitro development ability of the sperm was also examined with intracytoplasmic sperm injection (ICSI). The rate of male pronuclear formation was (68.52%), for cleavage (59.17%) and for blastocyst formation (19.16%) of the trehalose group (0.2 mol/L), significantly higher than those of the 50 mmol/L EDTA group (64.59%, 56.26% and 15.62%) and the control group (35.36%, 52.33% and 8.60%) (P < 0.05). After storage for 60, 120 and 180 d at 4 degrees C, no significant difference in the in vitro development was observed (P > 0.05). The male pronuclear, cleavage and blastocyst formation after ICSI with freeze-dried spermatozoa incubated for 1 h was superior than those incubated for 2 h (P < 0.05). No significant differences in the structures after stored at 4 degrees C or -20 degrees C (P > 0.05) were observed between the trehalose group and EDTA group. The percent of B grade freeze-dried boar spermatozoa in the trehalose group was higher than that of the EDTA group (P < 0.05). Based on the ultrastructure observation, main cryogenic damage in freeze-dried boar spermatozoa was swelling, damage or rupture in the sperm acrosome, neck and tail.
Animals
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Freeze Drying
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Male
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Semen Preservation
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methods
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veterinary
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Sperm Injections, Intracytoplasmic
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veterinary
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Spermatozoa
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Swine
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Trehalose
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pharmacology