Objective:To explore the improvement of the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection and conduct safety tests including abnormal toxicity test,allergic reaction test,hemolysis and coagulation test.Methods:Ginkgo Leaf Extract and Dipyridamole Injection from 3 different manufactures(A,B and C)were tested respectively through abnormal toxicity test and acute toxicity test in mice,active systemic anaphylaxis test in guinea pigs and hemolysis test in vitro.Five mice were used in each batch for abnormal toxicity test according to the abnormal toxicity test method in general notice of the Chinese Pharmacopoeia 2020 Volume Ⅳ(1141),and 50 mice were selected in each batch for acute toxicity test to determine the median lethal dose(LD50)or maximum tol-erable dose(MTD)of Ginkgo Leaf Extract and Dipyridamole Injection,which were used to establish the method of abnormal toxicity experiment.The anaphylaxis of Ginkgo Leaf Extract and Dipyridamole Injection was evaluated by active systemic anaphylaxis test in guinea pigs,which was used to establish the method of allergic test.The hemoly-sis test of Ginkgo Leaf Extract and Dipyridamole Injection was studied by conventional tube method in vitro(macro-scopic observation)and improved hemolysis method in vitro(spectrophotometric method),which were used to establish the method of hemolysis and coagulation test.Results:① In manufacture A,the results of abnormal toxicity test were showed that LD50 was20.8 mL·kg-1and MTD was 16.5 mL·kg-1.No death or abnormal reac-tions were observed in mice tested for abnormal toxicity of 2 manufactures(B and C),and MTD was 50 and 40 mL·kg-1,respectively.②The no-observed-adverse-effect dose of Ginkgo Leaf Extract and Dipyridamole Injec-tion from 3 manufactures to guinea pig intravenous was 0.83 mL·kg-1,and no allergic reaction symptoms were observed when Ginkgo Leaf Extract and Dipyridamole Injection was diluted 4 times to challenge the sensitized guinea pigs(equivalent to human clinical dosage).③Differences were observed in the hemolytic effects of Ginkgo Leaf Extract and Dipyridamole Injection from 3 manufactures,but no obvious hemolytic reaction occurred when it was diluted 1.2 times(equivalent to 5%of the maximum clinical concentration).Conclusion:It is recommended to add abnormal toxicity test,allergic reaction test,hemolysis and coagulation test in the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection as safety test items to control the risk.The proposed method is diluting Ginkgo Leaf Extract and Dipyridamole Injection by 5 times,4 times and 1.2 times to perform abnormal toxicity test,allergic reaction test,hemolysis test and coagulation test respectively.

Objective:To explore the improvement of the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection and conduct safety tests including abnormal toxicity test,allergic reaction test,hemolysis and coagulation test.Methods:Ginkgo Leaf Extract and Dipyridamole Injection from 3 different manufactures(A,B and C)were tested respectively through abnormal toxicity test and acute toxicity test in mice,active systemic anaphylaxis test in guinea pigs and hemolysis test in vitro.Five mice were used in each batch for abnormal toxicity test according to the abnormal toxicity test method in general notice of the Chinese Pharmacopoeia 2020 Volume Ⅳ(1141),and 50 mice were selected in each batch for acute toxicity test to determine the median lethal dose(LD50)or maximum tol-erable dose(MTD)of Ginkgo Leaf Extract and Dipyridamole Injection,which were used to establish the method of abnormal toxicity experiment.The anaphylaxis of Ginkgo Leaf Extract and Dipyridamole Injection was evaluated by active systemic anaphylaxis test in guinea pigs,which was used to establish the method of allergic test.The hemoly-sis test of Ginkgo Leaf Extract and Dipyridamole Injection was studied by conventional tube method in vitro(macro-scopic observation)and improved hemolysis method in vitro(spectrophotometric method),which were used to establish the method of hemolysis and coagulation test.Results:① In manufacture A,the results of abnormal toxicity test were showed that LD50 was20.8 mL·kg-1and MTD was 16.5 mL·kg-1.No death or abnormal reac-tions were observed in mice tested for abnormal toxicity of 2 manufactures(B and C),and MTD was 50 and 40 mL·kg-1,respectively.②The no-observed-adverse-effect dose of Ginkgo Leaf Extract and Dipyridamole Injec-tion from 3 manufactures to guinea pig intravenous was 0.83 mL·kg-1,and no allergic reaction symptoms were observed when Ginkgo Leaf Extract and Dipyridamole Injection was diluted 4 times to challenge the sensitized guinea pigs(equivalent to human clinical dosage).③Differences were observed in the hemolytic effects of Ginkgo Leaf Extract and Dipyridamole Injection from 3 manufactures,but no obvious hemolytic reaction occurred when it was diluted 1.2 times(equivalent to 5%of the maximum clinical concentration).Conclusion:It is recommended to add abnormal toxicity test,allergic reaction test,hemolysis and coagulation test in the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection as safety test items to control the risk.The proposed method is diluting Ginkgo Leaf Extract and Dipyridamole Injection by 5 times,4 times and 1.2 times to perform abnormal toxicity test,allergic reaction test,hemolysis test and coagulation test respectively.

Objective To establish a method for bacterial endotoxin test of flumazenil bulk drug.Methods According to the bacterial endotoxin test(BET)method in general rule 1143 in the Chinese Pharmacopoeia(2020 Edition,VolumeⅣ),flumazenil was dissolved with dimethyl sulfoxide(DMSO)and diluted with BET water.The gel-clot method and kinetic turbidimetric assay were used to carry out the interference test and endotoxin recovery test.Results Flumazenil was dissolved with DMSO to 10 mg·mL-1,and then diluted 100 times or more by BET water,which has no interference effects on the bacterial endotoxin test.Conclusion The BET method established in this study can be used to control the quality of flumazenil by performing bacterial endotoxin tests.

Objective: To observe the acute hypotensive effect of compound coenzyme for injection on cats,and to establish a method for examination of depressor substance.
Methods: Ten batches of compound coenzyme for injection and histamine depressor substance were compared by cat blood pressure method to determine the limit value of depressor substance test method. According to the limit value, 22 batches of samples were tested for depressor substance.
Results: The limit of compound coenzyme for injection was 3 IU·kg^-1 (calculated by coenzyme A). Two batches of 22 batches of compound coenzyme for injection did not meet the requirements.
Conclusion: The method of compound coenzyme for injection is feasible according to the proposed limit value. It is suggested that the quality standard of compound coenzyme for injection should be added with the examination of depressor substance.

OBJECTIVE：To provide reference for improving the operation efficiency of drug quality sampling and inspection in China . METHODS ：Starting from the application and management situation of inspection standards/methods in provincial inspection institutions ，the problems existing in the application and management of inspection standards/methods in provincial inspection institutions were analyzed ；the inspection standards/methods database of provincial inspection institutions is attempted to build，combining with the relevant experience and practices of FDA. RESULTS & CONCLUSIONS ：The inspection methods involved in drug sampling and inspection could be divided into official standards and non-standard methods. Official standards were the main standards for drug sampling and inspection ，and were mainly used for routine inspection. Such kind of standards could be classified according to the characteristics of compiled ，single-page and later-issued supplementary ；an electronic catalogue should be established for unified management. Non-standard methods were only used for sample preliminary screening ，verification of official inspection results ，quality evaluation and inspection of unknown or suspicious samples in emergency inspection. Its tracking，collection and management mechanisms were not yet complete. It is recommended to draw on the experience of drug sampling and inspection in the United States so as to establish method database hierarchically. For the mature method established in drug supervision system ，an electronic catalog and document content database should be established and the method should be confirmed before use ；for national standards and the recommended methods published by authoritative institutions in other industries，and mature methods published in scientific and technological literature ，the retrieval channels should be listed ，the methods should be verified ，reviewed and approved before use ，and an electronic catalogue should be established and recorded in time after use. The electronic catalogue format of non-standard methods generally include controlled number ，applicable variety name，method name ，inspection items ，etc.

Objective:To optimize water extraction and alcohol precipitation technology of Xiaoaiping capsules. Methods:With the content of Tenacissoside G and dry extract yield rate as the index, to investigate the effects of extraction times, extraction time and the amount of water on extraction technology by orthogonal test; and investigate the influence of the concentration of alcohol precipitation, the relatively density of concentrated liquid and storage time on alcohol precipitation technology by orthogonal test.Results:The optimal water extraction technology was 6-fold water, decocting 3 times for 1.5 h each time. The optimal alcohol precipitation technology was concentrating the water extraction to the density of 1.15 (60 ℃), and add alcohol after cooling with 70% alcohol standing for 12 hand then removing the precipitation.Conclusions:The optimized water extraction and alcohol precipitation process is stable, feasible and reproducible, which could provide reference for the extraction and purification of Xiaoaiping capsules.

BACKGROUND: Silk fibroin, the main component of silk fibroin biofilm, is a natural protein, but it is still a heterologous protein to the body. Its immunogenicity/toxicity is an important factor in determining the development prospects. OBJECTIVE: To evaluate the toxicity of absorbable silk fibroin biofilm and its degradation products on the rat immune system by muscle implantation experiments in rats. METHODS: Ten Wistar rats from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. were implanted with a long strip of 1 mm× 10 mm in the right gluteal muscle to observe the implant absorption and determine the implantation cycle. Seventy-two Wistar rats were assigned into control and experimental groups (n=36/group, 18 in either sexes) . A long strip of 1 mm×10 mm was implanted into the rat right gluteal muscle in the experimental group, and the control group received no implantation. Some of the rats were taken for histological examination and calculate the organ/weight ratio, at 26 weeks postoperatively. The spleen lymphocyte proliferation ability, NK cell activity, cell classification of T lymphocytes, and levels of interleukin 2 and tumor necrosis factor α were detected. Another part of the animals was taken for macrophage phagocytosis of erythrocytes cell capacity and antibody producing cell count. RESULTS AND CONCLUSION: (1) Silk fibroin biofilm was completely absorbed after implanted into the rat muscle for 26 weeks. (2) In female or male Wistar rats, the immune organs in the experimental group showed no significant changes in the appearance, weight and histological examination. There were no significant differences in the hematological indexes (hemoglobin, red blood cell count, hematocrit, blood platelet count and white blood cell count and classification) , spleen lymphocyte proliferation, NK cell activity, the ratio of T lymphocytes and their subpopulations, and the levels of interleukin 2 and tumor necrosis factor α had no significant differences between two groups (P> 0.05) . (3) In female or male Wistar rats, the macrophage phagocytosis of chicken erythrocytes cell capacity and antibody producing cell count showed no significant differences between two groups (P> 0.05) . (4) These results indicate that silk fibroin biofilm causes no immunosuppression or immunostimulation on immune organs, immune cells and immune molecules (cytokines) of rats.

Objective To establish and validate the Caco-2 cell in vitro absorption model,so as to play a foundation for next study of drug absorption and transport. Methods Caco-2 cells were cultured on the polycarbonate membrane inserts fixed in the 24-well transwell transport chamber with three types cell density of low,medium and high concentration (5×104,1×105,2× 105·mL-1) respectively.After being cultured for 3,6,9,12,15,18,21 d,the integrity of monolayer were assessed and compared by the cell morphology,growth characters and transepithelial electrical resistance (TEER),aiming to determine the inoculum density and culture time.Then the permeability and polarity were validated by the apical-to-basolateral amount of Lucifer Yellow across cell monolayer,the alkaline phosphatase activity in the apical side (AP),the basolateral side (BL) and intracellular activity. Results The cells of low,medium and high concentration group had fusion into a integrate cell monolayer and the maximum absorbance after being cultured for 15,12,9 d respectively.However,conglobated and dead cells were observed at the later growth stage in the medium and high concentration group and the TEER of cell monolayer were smaller than the low concentration group,which could reach 300 Ω·cm2after cultured 15 d and keep a relatively stable value, then cells were cultured with 5×104·mL-1cell density for 21 d.The Lucifer Yellow apparent permeability coefficient (Papp) was 3.57×10-7 cm·s-1which was lower than 5. 0×10-7cm·s-1according to provision. And the intracellular alkaline phosphates’ activity increased,AP/BL increased by 5 times in day 21. Conclusion The integrity,permeability and polarity of the established Caco-2 cell model in our laboratory was validated,and it can be used as an in vitro model similar with small intestinal epithelium for absorption and transport studies.

Objective:To study the acute toxicity in mice and cytotoxicity of the impuritiy(3-amino-2-piperidinone hydrochloride) in ornithine. Methods:The mice were given the impurity by intravenous injection.The acute toxicity was observed and LD50in mice was calculated by Bliss method. According to the result of LD50,the mice were given the impurity respectively at high,medium and low dose. The changes of general condition and body weight were recorded,and the blood biochemical indices and histomorphology of organs and tissues were observed after 14 days. The cytotoxicity was measured on fibroblasts(L929) cells. Results:The LD50and 95% con-fidence limit of the impurity (intragastric administration) was 758.49-848.08 mg·kg-1.All the detected indices of the three groups (660,330 and 170 mg·kg-1) including general condition,weight change,biochemical indices,organ morphological and histological change were all within the normal ranges. The impurity significantly decreased the viability of L929 cells. The NOAEL value was 660 mg·kg-1in mice. Conclusion:The impurity has certain toxicity to mice and cells. The data can provide experimental basis for the quality standard establishment for ornithine and the safety improvement in clinic use.

Objective:To study the acute toxicity and genotoxicity of Sanqi liver-protecting capsules to evaluate the toxicological safety. Methods:The mouse acute oral toxicity test, Ames test, micronucleus test of mouse bone marrow and mouse sperm shape ab-normality test were carried out for Sanqi liver-protecting capsules. Results:The mouse acute oral maximum tolerance dose ( MTD) of Sanqi liver-protecting capsules was above 20 g·kg-1 in the mouse acute oral toxicity test, which showed a non-toxic substance. The results of Ames test, micronucleus test of mouse bone marrow and mouse sperm shape abnormality test were negative in all the groups. Conclusion:Under the above experimental conditions,the genotoxicity of Sanqi liver-protecting capsules is not found out, therefore, the capsules are classified as a non-toxic drug.