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.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
3.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
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.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
7.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
8.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
9.Freeze grinding combined ultrasonic technique for the treatment of nail test material.
Hang CHEN ; Ping XIANG ; Qi-ran SUN ; Min SHEN
Journal of Forensic Medicine 2012;28(5):342-346
OBJECTIVE:
To investigate the feasibility of the new method of combining freeze grinding with ultrasonic technique for the pretreatment of the nail for toxicological and pharmaceutical analysis and to compare the advantages and disadvantages of this method with other traditional methods.
METHODS:
Five pretreatment methods were examined. Scanning electron microscope (SEM) was used to observe the microstructural changes of the nail.
RESULTS:
The microscopic structure of nail totally destroyed after alkali treatment. The hierarchy mode of the internal structure became obvious and tight after acid hydrolysis, which became indistinct after methanol infiltration. The structure of nail broke to pieces after ultrasonic technique. After freeze grinding combined ultrasonic technique, the particle structure kept original shape, and its size was one hundred times smaller than which after manual way.
CONCLUSION
The freeze grinding combined ultrasonic technique can improve the release efficiency, and ensure the stability of the toxicant or drug during the pretreatment process. It is appropriate for toxicological and pharmaceutical analysis in the nail.
Forensic Pathology
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Freeze Drying/methods*
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Humans
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Microscopy, Electron, Scanning
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Nails/chemistry*
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Particle Size
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Ultrasonics
10.Microencapsulated β-carotene preparation using different drying treatments.
Xiang-Yu LI ; Mian-Bin WU ; Min XIAO ; Shu-Huan LU ; Zhi-Ming WANG ; Jian-Ming YAO ; Li-Rong YANG
Journal of Zhejiang University. Science. B 2019;20(11):901-909
β-Carotene is one of the most abundant natural pigments in foods; however, usage of β-carotene is limited because of its instability. Microencapsulation techniques are usually applied to protect microencapsulated β-carotene from oxidization. In this study, β-carotene was microencapsulated using different drying processes: spray-drying, spray freeze-drying, coating, and spray granulation. The properties of morphology, particle size, water content, thermal characteristic, and chemical stability have been explored and compared. Scanning electron microscopy measurements showed that the coated powder had a dense surface surrounded by starch and suggested that the coating process gave a microencapsulated powder with the smallest bulk density and the best compressibility among the prepared powders. The chemical stabilities of microcapsules were evaluated during six months of storage at different temperatures. The coated powder had the highest mass fraction of β-carotene, which indicated that the coating process was superior to the three other drying processes.
Drug Compounding/methods*
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Drug Stability
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Freeze Drying
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Microscopy, Electron, Scanning
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Technology, Pharmaceutical
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beta Carotene/chemistry*