Aszalos A, Crawford J, Vollmer P, Kantor N, Alexander T
J Pharm Sci. 1981 Aug;70(8):878-80. doi: 10.1002/jps.2600700811.
A fast and sensitive method was developed for the quantitative determination of at least 10 components of pharmaceutical bleomycin sulfate preparations. The method is based on the reversed-phase high-performance liquid chromatographic (HPLC) separation of the components on a muBondapak C18 column with a mobile phase having a linear gradient of 10--40% methanol in aqueous 0.005 M 1-pentanesulfonic acid at pH 4.3. With this assay, the average standard deviations for components A2 and B2 are 0.92 and 0.87, respectively, for a 7.5--22.5 x 10(-3)-mg sample. Regulatory agencies presently use the official Code of Federal Regulations (CFR) method, which is based on CM-Sephadex column chromatography. It was demonstrated that this CFR method does not separate the bleomycin A2 component from some other minor bleomycin components. After elution from the CM-Sephadex column, the "A2 component" was separated into five components by the HPLC method. Bleomycin A2 is stable under these HPLC conditions.
已开发出一种快速灵敏的方法，用于定量测定硫酸博来霉素制剂中的至少10种成分。该方法基于在μBondapak C18柱上对各成分进行反相高效液相色谱（HPLC）分离，流动相为pH 4.3的0.005 M 1-戊烷磺酸水溶液中甲醇含量呈10% - 40%线性梯度的溶液。采用该测定法，对于7.5 - 22.5×10⁻³ mg的样品，成分A2和B2的平均标准偏差分别为0.92和0.87。监管机构目前使用基于CM - 葡聚糖凝胶柱色谱的官方联邦法规（CFR）方法。结果表明，该CFR方法无法将博来霉素A2成分与其他一些次要的博来霉素成分分离。从CM - 葡聚糖凝胶柱洗脱后，通过HPLC方法将“A2成分”分离为五种成分。博来霉素A2在这些HPLC条件下稳定。
