ABSTRACT
Objective To optimize the detection test of Pig-a gene mutation in peripheral blood of rats by enriching and detecting mutant erythrocytes, using immunomagnetic separation technique in combination with flow cytometry. Methods SD rats were administered with 20,40 and 80 mg/(kg·bw)doses of N-ethyl-N-nitrosourea(ENU) continually for 3 days. The peripheral blood samples of rats were collected on the 7th,14th and 28th days, respectively, after treatment. Immunomagnetic separation columns were used to enrich RETCD59- and RBCCD59- cells, and then flow cytometry was used to count the number of pre-column and post-column peripheral erythrocytes. Results Compared with the control group,the frequencies of RETCD59- and RBCCD59- were significantly increased in each ENU group(P<0.05). With immunomagnetic separation technique, the test of Pig-a gene mutation of a sample could be completed within 3 minutes,and the number of detected RETCD59- or RBCCD59-cells was up to 2×104or 9×104, respectively. Conclusions In this study,immunomagnetic separation in combination with flow cytometry is used to establish and optimize the Pig-a gene mutation test in rat peripheral blood,showing a high-throughput detection and improved accuracy and efficiency of the experiment.
ABSTRACT
Objective:To establish the test method for histamine-like substances in pirarubicin. Methods:According to the meth-od described in the appendix of Chinese Pharmacopoeia (2010 edition), the test for histamine-like substances was carried out. Re-sults:The test for histamine-like substances had no interference under the condition with pirarubicin concentration of 0. 01mg·ml-1 . The established test method for histamine-like substances in pirarubicin was applied in 10 samples, and the results were accordant with those of the test method for depressor substances. Conclusion:The test method for histamine-like substances in pirarubicin is feasible with the limit of 1 μg·mg-1 .
ABSTRACT
Skin sensitization (allergic contact dermatitis, ACD), is a serious condition caused by small reactive molecules and is characterized by a delayed-type hypersensitivity .Animal tests were usually used in the evaluation of sensitizing potential of chemical substances in the past .However , there is an increasing interest from the public for reducing and ultimately replacing animal tests .The European Union (EU) has posed a ban on animal testing of cosmetic ingredients that includes skin sensitization since 2013.Therefore, alternative in vitro tests are the main tendency in chemical substances and cosmetic sensitizing potential research in the future .In this study, different kinds of in vitro test methods that were adopted by OECD or on research (LLNA, DPRA, KeratinoSens TM, h-CLAT) were reviewed through recent years literature , comprehensive introduction and evaluation were made to obtain reliable hazard and potency information on potential skin sensitizers .
ABSTRACT
Objective:To establish a test method for histamine-like substances in aclarubicin hydrochloride. Methods:According to the method in the appendix of Chinese Pharmacopoeia ( edition 2010 ) , the test of histamine-like substances was carried out. Re-sults:The test of histamine-like substances was not interfered by aclarubicin hydrochloride with the concentration below 0. 01 mg· ml-1 . Histamine-like substances in 4 batches of samples were determined by the established method, and the results were consistent with those of the test method for depressor substances. Conclusion: The test method for histamine-like substances is feasible with the limit of 1 μg·mg-1 .