| Objective:Our group study the optimum required pH value of fresh Vibrio fischeri to determine the optimum growth pH of Vibrio fischeri,and theinitial luminous intensity and decay rate of the cμltured Vibrio fischeri reached the laboratory test requirements.Meanwhile,using the Microtox test technology,we tested the toxicity of traditional Chinese medicine injections reported by the National Adverse Reaction Monitoring Center and 9kind of Chinese traditional medicine injections provided by several pharmaceutical companies.Methods:According to ISO-11348 the fresh bacterial preparation methods,the growth of Vibrio fischeri was investigated at pH 4,5,6,7,8.After determining the optimal pH of Vibrio fischeri,the stability of freshly prepared Vibrio fischeri was investigated and the methodology was investigated(repeatability and precision).Based on the optimal detection system of microtox assay of TCM injections established by our laboratory and freshly prepared Vibrio fischeri,we evaluated the luminous biological effect and quality control of 9 batches of TCM injections from 8 manufacturers and its vehicle.Results:When pH was4～7,the growth of the bacteria was the best,suitable for the follow-up test.The resμlts of freshly prepared Vibrio fischeri showed that theluminescence intensity of Vibrio fischeri decreased over time,then tended to be stable state from 15 to 60 minutes.Under the optimal testing system established by laboratory,the relative standard deviations of repeatability test and intermediate precision test were both less than 15%.The test resμlts of 27 batches of 9 samples of TCM injections and three batches of the test resμlts of the vehicle showed that the IC50 values of three batches of C injection were(29.59± 0.02)%,41.29±7.02 36.56± 1.94)%;The IC50 values of three batches of injection were(3.72± 0.12,4.44±1.00%,4.26 ±1.05);The IC50 values of three batches of D injection were(4.26 ± 3.29,4.36 ±1.93,4.36± 1.64)%,;E injection(3.98±1.18,2.48 ± 0.71,4.33±0.49)%there was not statistically significant different;IC50 values of three batches of F injection(1.01± 0.15,1.36± 0.16,1.18±0.26)%;IC50 values of three batches of G injection(3.76±0.60,3.88± 0.01,3.51±0.18)%,IC50 values of three batches of I was(1.74± 0.56,1.74± 0.21,1.68± 0.18)%,IC50 values of three batches of H injection was(10.55± 3.43,10.82± 3.62,9.18 ± 0.99);The IC50 values of three batches of A injection solution were(5.60%±0.39%),(5.42 ± 0.22%),(5.59 ± 0.10%),there was no significant difference between batches(P>0.05).The IC50 values of seven batches of A injection were(2.25 ±0.46)%,(3.77±0.65)%,(3.56±0.55)%,(2.49±0.40)%,(2.73±0.30)%,2.95± 0.55)%,(1.39± 0.11)%respectively.There was significant difference between different batches(p<0.05 or 0.01).Conclusions:The optimal pH of laboratory cμltured Vibrio fischeri is 7.The initial luminous intensity and decay rate of freshly prepared Vibrio fischeri meet the laboratory standards for testing the biological toxicity of TCM injections.Using freshly cμltured Vibrio fischeri to carry out quality control tests on different batches of samples sent from different manufacturers showed that there was a significant concentration-effect of the injection injection on the luminescent biological effects of Vibrio fischeri between batches of injections of the same manufacturer relationship,the IC50 valueof differencedifferent batches ofthe same injection was not statistically significant.The IC50 value difference of different batches of same injection from different manufacturers was statistically significant(p<0.05).Prompt,1)Microtox technology can be used to test for the combined bio-effect(toxicity)detection of injections.2)On the other hand,the injection products of the same manufacturers possess good consistency.In further research,the number of test samples shoμld be expanded to establish the upper and lower limit of micro-toxic biological effects of injection quality control,which can help to further control the quality of injection.3)A injection had significant concentration-effect relationship on the luminescence of Vibrio fischeri,and the IC50 values of some batches were significantly different.Tip:Microtox technology can not only be used to detect A synthetic biological effects,but also for product quality control also has good prospects. |
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