HPLC法测定琥珀酰明胶注射液中的游离琥珀酸及琥珀酰化度

目的: 建立测定琥珀酰明胶注射液中游离琥珀酸及琥珀酰化度的高效液相色谱法. 方法: 采用Agilent Zorbax 300SB-C₁₈色谱柱(4.6 mm×250 mm,5 μm),流动相为乙腈-0.02 mol·L^-1磷酸二氢钾缓冲液(pH 2)(5:95),流速0.6 mL·min^-1,柱温20 ℃,检测波长210 nm. 结果: 分离度良好,满足定量测定要求,线性范围为30~480 μg·mL^-1,加样平均回收率均大于90%,琥珀酰明胶注射液样品中游离琥珀酸含量小于200 mg·L^-1,琥珀酰化度为1.9%,均符合要求. 结论: 建立的方法简便快速,重复性好,精密度高,可用于测定琥珀酰明胶注射液中游离琥珀酸及琥珀酰化度.

Objective: To establish an HPLC method for the determination of free succinylated gelatin and the succinylation degree in succinylated gelatin injection. Methods: The Agilent Zorbax 300SB-C₁₈ column(4.6 mm×250 mm,5 μm)was used with acetonitrile-0.02 mol·L^-1 KH₂PO₄ buffer solution(pH 2)(5:95)as the mobile phase,with the flow rate of 0.6 mL·min^-1,the column temperature of 20 ℃,and the detection wavelength of 210 nm. Results: In the range of 30-480 μg·mL^-1,the linearity relationship of succinic acid was good.The accuracy and repeatability was high.The average recovery rate of succinic acid was above 90%.The content of free succinylated gelatin was lower than 200 mg·L^-1 and the succinylation degree was 1.9%,which met the requirements. Conclusion: The method with a single column is rapid,repeatable,precise and convenient,which can be used for the determination of free succinic acid and succinylation degree in succinylated gelatin injection.

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