高效液相色谱法测定生长抑素中4种特定杂质

Determination of the Four Specified Impurities of Somatostatin by HPLC

目的:本文建立高效液相色谱法测定生长抑素特定杂质的方法。方法:采用十八烷基硅烷键合硅胶为填充剂(Agilent ZORBAX SB-C18, 4.6 mm ×250 mm, 3.5 μm);以乙腈为流动相A,以0.05 mol∙L−1六氟磷酸钾,0.02 mol∙L−1磷酸氢二铵水溶液(磷酸调pH3.5)为流动相B,梯度洗脱;流速为每分钟1.0 mL;检测波长为220 nm;柱温为30℃;进样体积15 μL。结果:生长抑素与相邻杂质峰(Di-Gly2-生长抑素、Des-Gly2-生长抑素)的分离良好;生长抑素氧化杂质1、生长抑素氧化杂质2、Di-Gly2-生长抑素和Des-Gly2-生长抑素在浓度范围内呈现良好的线性关系(相关系数 ≥ 0.9996);各杂质回收率均在80%~120%范围内,RSD ≤ 3.22%。结论:本法可用于生长抑素中特定杂质的检测。

Objective: To develop an HPLC method for the determination the known impurities of Somatostatin. Methods: The analysis was conducted on octadecylsilane-bonded silica gel (Agilent ZORBAX SB-C18, 4.6 mm ×250 mm, 3.5 μm);The mobile phase A: Acetonitrile;The mobile phase B: phosphate buffer (0.05 mol∙L−1 potassium hexafluorophosphate and 0.02 mol∙L−1 diammonium hydrogen phosphate, adjust the pH to 3.5 with phosphoric acid) with elution at the flow rate of 1.0 mL∙min−1;The column temperature was 30˚C, the detection wavelength was 220 nm and the injection volume was 15 μL. Results: The known impurities (Di-Gly2-somatostatin and Des-Gly2-somatostatin) were completely separated from somatostatin. The results showed that there was a good linear relationship (correlation coefficient ≥ 0.9996) in the concentration range of somatostatin oxidation impurity 1, somatostatin oxidation impurity 2, Di-Gly2-somatostatin and Des-Gly2-somatostatin in the concentration range (correlation coefficient ≥ 0.9996);the recovery of each impurity was in the range of 80%~120%, RSD ≤ 3.22%. Conclusion: The established method is suitable for the determination of the known impurities of somatostatin.