Buck R H, Maxl F
Analytical Research and Development and Quality Assurance, Sandoz Pharma Ltd. Basle, Switzerland.
J Pharm Biomed Anal. 1990;8(8-12):761-9. doi: 10.1016/0731-7085(90)80118-9.
A high-performance liquid chromatographic (HPLC) method is described for the assay of salmon calcitonin. The method uses a 5-microns octadecasilyl silica column (100 x 4.6 mm) at 50 degrees C and an initial mobile phase (flow rate 1 ml min-1) comprising 35% of B (1 M tetramethylammonium hydroxide-water-acetonitrile, 8:392:600) and 65% of A (1 M tetramethylammonium hydroxide-water-acetonitrile, 20:880:100) with linear gradient elution over 21 min to a final mobile phase of 57% B; solutions A and B are adjusted to pH 2.5 with phosphoric acid. Detection was by UV spectrophotometry at 210 nm. The method has been shown to be selective, precise and rapid and, in a collaborative study to give excellent correlation with the results obtained by using the biological assay method of the European Pharmacopoeia. The method, which has been applied successfully to the assay of different batches of salmon calcitonin in bulk drug and in formulated products, is recommended for adoption as the pharmacopoeial assay method.
本文描述了一种用于测定鲑鱼降钙素的高效液相色谱（HPLC）方法。该方法使用一根5微米的十八烷基硅烷硅胶柱（100×4.6毫米），柱温50℃，初始流动相（流速1毫升/分钟）由35%的B（1M四甲基氢氧化铵-水-乙腈，8:392:600）和65%的A（1M四甲基氢氧化铵-水-乙腈，20:880:100）组成，在21分钟内进行线性梯度洗脱至最终流动相为57%B；溶液A和B用磷酸调节至pH 2.5。检测采用210nm紫外分光光度法。该方法已被证明具有选择性、精密度高且快速，并且在一项协作研究中，与采用欧洲药典生物测定法获得的结果具有良好的相关性。该方法已成功应用于原料药和制剂中不同批次鲑鱼降钙素的测定，建议采用该方法作为药典测定方法。
