RP-HPLC法测定盐酸氯胺酮注射液有关物质和含量

目的：通过对现有质量标准中有关物质检查HPLC法色谱条件的优化，建立新的盐酸氯胺酮注射液有关物质和含量测定用HPLC方法，改善现有色谱体系拖尾问题并提高杂质的检出率。 方法：以十八烷基硅烷键合硅胶为填充剂，以 0.0025 mol·L^-1庚烷磺酸钠（0.2%三乙胺，0.08%醋酸）-乙腈（50：50）为流动相，流速1 mL·min^-1，检测波长215 nm，柱温30℃，进样体积20 μL。 结果：新建色谱条件能够使氯胺酮与其已知杂质羟亚胺达到完全分离；盐酸氯胺酮浓度在0.001~1 mg·mL^-1范围内呈良好的线性关系（r=0.998）；氯胺酮的检测限（S/N=3）和定量限（S/N=10）分别为1.5 ng和5 ng，羟亚胺的检测限（S/N=3）和定量限（S/N=10）为5.9 ng和19.8 ng；该方法精密度良好；加样回收率（n=9）为98.69%；盐酸氯胺酮及羟亚胺在乙腈-水（4：1）溶液中24 h内稳定。 结论：新建色谱条件在改善氯胺酮峰形的同时提高了方法的专属性和准确性，适用于盐酸氯胺酮注射液中有关物质检查和含量测定。

Objective: To establish a new method for related substance detection and content determination of ketamine hydrochloride injection through the optimization of the existing HPLC chromatogram conditions,so as to solve the tailing problem of chromatographic system and improve the detection rate of impurities of current official pharmacopeia. Methods: This new method was carried out on a Chromolith RP-C₁₈column using a mixture of 0.0025 mol·L^-1 sodium heptanesulfonate buffer(0.2% triethylamine,0.08% glacial acetic acid) and acetonitrile (50: 50,v/v) at a flow rate of 1 mL· min^-1;the detection was done at the wavelength of 215 nm,the column temperature was 30℃,and the injection volume was 20 μL. Results: A good resolution with baseline separation between ketamine and the well-known impurities hydroxylimine was obtained.The LOD(S/N=3) and LOQ(S/N=10) of ketamine were 1.5 ng and 5 ng,respectively;The LOD(S/N=3) and LOQ(S/N=10) of hydroxylimine were 5.9 ng and 19.8 ng,respectively.Standard curves were linear over the range of 0.001 to 1 mg·mL^-1 with all correlation coefficient values being 0.998.The average recovery rate(n=9) was 98.69%. Ketamine hydrochloride and hydroxylimide were stable in acetonitrile water (4: 1) for at least 24 h. Conclusion: Compared with the original method,this new method is accurate,simple and specific,which is suitable for related substance detection and content determination of ketamine hydrochloride injections.

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