目的 建立测定鲑降钙素鼻用喷雾剂中有关物质的HPLC方法。方法 使用Macherey-Nagel Nucleosil 100-5 C₁₈AB色谱柱（250 mm×4.0 mm，5 μm）；以0.04 mol/L四甲基氢氧化铵溶液（用磷酸调pH 2.5）–乙腈（9∶1）为流动相A，以0.04 mol/L四甲基氢氧化铵溶液（用磷酸调pH 2.5）–乙腈（4∶6）为流动相B，梯度洗脱；检测波长为220 nm；体积流量为1.0 mL/min；进样量为100 μL；柱温为65 ℃；样品室温度：10 ℃。按加校正因子的主成分自身对照法计算降钙素C、N-乙酰半胱氨酰鲑降钙素（杂质A）、9-D-亮氨酸鲑降钙素（杂质B）、去-22-酪氨酸鲑降钙素（杂质C）。结果 降钙素C和鲑降钙素杂质A、B、C在0.20～7.50 μg/mL线性关系良好，平均回收率分别为98.1%、98.9%、100.5%、99.7%，RSD值分别为1.6%、1.5%、1.0%、1.1%。杂质A、B、C的校正因子均超出0.90～1.10。结论 方法简便、快速，可作为鲑降钙素鼻用喷雾剂中有关物质的检测方法。

Objective To establish an HPLC method for determination of related substance in Salmon Calcitonin Nasal Spray. Methods The separation was performed on Macherey-Nagel Nucleosil 100-5 C₁₈AB column (250 mm × 4.0 mm, 5 μm). The mobile phase A was 0.04 mol/L tetramethyl ammonium hydroxide buffer (adjusted with phosphoric acid to pH 2.5) - acetonitrile (9∶1), and the mobile phase B was 0.04 mol/L tetramethyl ammonium hydroxide buffer (adjusted with phosphoric acid to pH 2.5) - acetonitrile (4∶6), with gradient elution. The detection wavelength was 220 nm, the flow rate was 1.0 mL/min, injection volume was 100 μL, column temperature was 65 ℃, and sample temperature was 10 ℃. The contents of calcitonin C and N-acetylcysteine salmon calcitonin (impurity A), 9-D-leucine salmon calcitonin (impurity B), and de-22-tyrosine salmon calcitonin (impurity C) were determined by principal component self-control with correction factor. Results The linear ranges of calcitonin C and impurity A, B, and C were all 0.20 - 7.50 μg/mL. The average recoveries were 98.1%, 98.9%, 100.5%, and 99.7% with RSD values of 1.6%, 1.5%, 1.0%, and 1.1%, respectively. The correction factors of impurity A, B, and C were all above 0.90 - 1.10. Conclusion The method is simple, rapid, and can be used for the determination of related substances in Salmon Calcitonin Nasal Spray.

R927.1