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OBJECTIVE:To adopt gel method for the determination of bacterial endotoxin in Fat emulsion(10%)/amino acid (15)/glucose (20%) injection. METHODS:According to the gel method in term ofbacterial endotoxin test methodin Chinese Pharmacopeia(2015 edition),the maximal valid dilution(MVD)of samples were determined through interference test and the vali-dated. The results were compared with chromogenic method. RESULTS:In gel method,the interference to agglutination reaction of TAL and bacterial endotoxin can be excluded when samples were diluted 24 times or less. In chromogenic method,the samples should be diluted 76 times or less. CONCLUSIONS:Gel method can be used for bacterial endotoxin test of Fat emulsion(10%)/amino acid(15)/glucose(20%)injection.
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OBJECTIVE:To establish the detection method for pyrogen-related factor interleukin 1β(IL-1β)through optimiz-ing detection condition,and to conduct preliminary methodology study. METHODS:The in vitro fresh human whole blood detec-tion method was used. The bacterial endotoxin standard solution(5,2,0.8,0.32 EU/ml)were added into diluted blood;using di-luted RPMI 1640 as blank control,the content of IL-1β in blood sample was determined by ELISA after incubation. The relation-ship of the addition of different attenuants(RPMI 1640 culture,sterilized normal saline)and fetal bovine serum,final dilution vol-ume fraction(40%,20%,10%and 8.3%)and storage duration(2,5,6,8,26 h)with the contents of endotoxin IL-1βwere in-vestigated,and related coefficient and detection limits were calculated. Different dilution times of Qingkailing injection and Ginaton injection samples and interference solutions were added into diluted blood to detect their recovery. RESULTS:The results indicated that RPMI 1640 media and 40% diluted blood was more sensitive(detection limit was 0.128 EU/ml,r=0.993);while the addition of fetal bovine serum didn’t influence the results. The detection limits of blood sample storied at 4 ℃ for 26 h were 0.128 EU/ml (r>0.990). When Qingkailing injection and Ginaton injection were diluted 10,32 and more times,the detection method was not interfered and the recovery ranged 68%-118%. CONCLUSIONS:Established in vitro fresh human whole blood detection method can be used for the detection of pyrogen,and provides trial evidence for the pyrogen detection of TCM injection.
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When some testing institutions performed biological evaluation to the disposable medical syringe piston, cytotoxicity was found. According to the biological evaluation testing Selection Guide proposed by Ministry of Health and the Comments of Sample Provider, We performed biological evaluation to one sample by using 5 tests of basic biological evaluation. Cytotoxicity was found, which was probably caused by the residue of the lotion. This research provides reference for objective evaluation of disposable medical syringe piston and safe guarantee of the product.
Assuntos
Testes Imunológicos de Citotoxicidade , Equipamentos Descartáveis , SeringasRESUMO
OBJECTIVE: To establish bacterial endotoxin test for adenosine disodium triphosphate (ATP) raw material by TAL method. METHODS: The sensitivity of TAL and the limit of endotoxin were reviewed and confirmed. Preliminary interference test was used to determine the dilution ratio of samples and interference test was applied to determine maximum non-interference concentration. Bacterial endotoxin test was performed on 11 batches of samples according to the method stated in Chinese Pharmacopoeia (2005 edition). RESULTS: The sensitivity of TAL was up to the standard and the limit of endotoxin in samples were 2.0 EU?mg-1. The maximum non-interference concentration was 0.125 mg?mL-1. Results of bacterial endotoxins test of 11 batches of samples were all in line with the standard. CONCLUSIONS: It is feasible to detect the bacterial endotoxin of ATP raw material by TAL methods, which can replace pyrogen test.