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盐霉素是第一个用于预防和治疗鸡球虫病的神经元钾通道开放剂.采用HPLC-UV、LC-MS等方法对盐霉素原料药中相关杂质进行检测.通过柱后衍生化高效液相色谱法检测到盐霉素原料药中共含有10个含量在0.18％-3.47％之间的杂质,其中3个杂质含量在2.0％以上.含量为2.04％的杂质为已知的20-氧代-盐霉素,其由盐霉素C-20位的仲羟基氧化为羰基而形成,但其他2个杂质(SAL-A含量为3.47％,SAL-B含量为2.30％)未见文献报道.通过制备液相色谱(Prep LC)从盐霉素原料药中分离得到该新杂质,并利用红外吸收光谱(IR)、一维及二维核磁(NMR)、高分辨质谱(HR-ESI-MS)和圆二色谱(CD)等测试分析方法对其化学结构进行表征,对这些表征数据进行综合解析,确证了2个盐霉素杂质(SAL-A和SAL-B)的化学结构:SAL-A为盐霉素C-6位的甲基变换为了乙基,SAL-B为盐霉素C-14位的甲基变换为了乙基.SAL-A和SAL-B的CD光谱数据也同盐霉素相似,说明它们具有相同的相对构型.此外,生物活性研究表明SAL-A和SAL-B对人肺癌细胞A549、人肝癌细胞HepG2和乳腺癌细胞SK-BR3具有一定的抑制活性.上述结果表明两个可作为潜在抗肿瘤药物的盐霉素新杂质得到发现和确证.“,”Salinomycin was the first neuronal potassium channel opener for the prevention and treatment of coccidiosis in poultry.The identification of related impurities of the salinomycin bulk drug was detected by HPLC-UV and LC-MS.Ten impurities ranging from 0.18％to 3.47％in the salinomycin bulk drug were detected by HPLC with post-column derivatization,and three impurities were present at levels greater than 2.0％.One impurity,20-oxo-salinomycin(2.04％),was a known compound that occurred as a result of oxidation of a secondary alcohol group to a ketone at the C-20 position,and the other two impurities(SAL-A present at 3.47％,and SAL-B present at 2.30％)were unknown and have not been reported previously.The two unknown impurities were obtained from the salinomycin bulk drug sample by preparative liquid chromatography(Prep LC).A thorough study was undertaken to characterize these two impurities using IR,1D(1H,13C,H-D,DEPT 90 and 135),2D(COSY,HSQC,HMBC)NMR,HR-ESI-MS,and CD spectral data.Based on the characterization data,SAL-A presented an ethyl group rather than a methyl group at the C-6 position of salinomycin,while SAL-B contained an ethyl group instead of a methyl group at the C-14 position of salinomycin.Similar CD spectral data of SAL-A and SAL-B to salinomycin indicated that their relative configurations were identical.The anti-tumor activity against human tumor cell liver HepG2,breast SK-BR3,and the non-small cell lung cancer cell line A549,revealed their ability to inhibit the growth of tumor cells.Therefore,two new impurities,SAL-A and SAL-B,in the salinomycin bulk drug at relatively low levels,were detected,isolated,and identified.Moreover,SAL-A and SAL-B showed anti-tumor activities against HepG2,SK-BR3,and A549 cells.