土霉素原料及片剂中有关物质测定  

Determination of Related Substances in Oxytetracycline Active Pharmaceutical Ingredients and Oxytetracycline Tablets

作　　者：杨天风 张少萌 高燕霞 张冬 YANG Tianfeng;ZHANG Shaomeng;GAO Yanxia;ZHANG Dong(Hebei Institute of Drug and Medical Device Inspection,Shijiazhuang,Hebei,China 050200;Hebei Drug Evaluation Center,Shijiazhuang,Hebei,China 050011)

机构地区：[1]河北省药品医疗器械检验研究院,河北石家庄050200 [2]河北省药品审评中心,河北石家庄050011

出　　处：《中国药业》2024年第5期68-72,共5页China Pharmaceuticals

基　　金：河北省科学技术厅中央引导地方科技发展资金项目[216Z4802G]。

摘　　要：目的建立测定土霉素原料及片剂中有关物质含量的高效液相色谱法。方法色谱柱为Thermo Hypersil GOLD Sino Chrom C_(18)柱(250 mm×4.6 mm,5μm),流动相A为醋酸铵溶液[0.25 mol/L醋酸铵-0.05 mol/L乙二胺四乙酸二钠溶液-三乙胺(100∶10∶1,V/V/V),用醋酸调pH至7.5]-乙腈(88∶12,V/V),流动相B为乙腈进行梯度洗脱,流速为1.0 mL/min,检测波长为254 nm,柱温为30℃,进样量为10μL。结果土霉素的质量浓度在0.5~20μg/mL范围内与峰面积线性关系良好(r=0.9998,n=6);检测限为1.4 ng,定量限为5.0 ng;精密度和稳定性试验结果的RSD均小于2.0%;专属性试验与强力破坏试验中,土霉素与强制破坏的降解产物及与其他8种已知杂质对照品均可实现有效分离。土霉素片中2-乙酰-2-去酰胺土霉素检出量为1.62%~2.73%,其他杂质总含量为1.20%~1.84%。结论该方法操作简便、专属性好、灵敏度高,可用于土霉素原料及片剂中有关物质的测定。拟订土霉素片中有关物质2-乙酰-2-去酰胺土霉素的限度为不得过对照溶液主峰面积的3.5%,其他各杂质峰面积之和不得过对照溶液主峰面积的2.0%。Objective To establish a high-performance liquid chromatography(HPLC)method for the content determination of related substances in oxytetracycline active pharmaceutical ingredients(API)and Oxytetracycline Tablets.Methods The chromatographic column was Thermo Hypersil GOLD SinoChrome C18 column(250 mm×4.6 mm,5μm),the mobile phase A was ammonium acetate solution[0.25 mol/L ammonium acetate-0.05 mol/L disodium ethylenediaminetetraacetate solution-triethylamine(100∶10∶1,V/V/V),pH was adjusted to 7.5 with acetic acid]-acetonitrile(88∶12,V/V),the mobile phase B was acetonitrile with gradient elution,the flow rate was 1.0 mL/min,the detection wavelength was 254 nm,the column temperature was 30℃,and the injection volume was 10μL.Results The linear range of oxytetracycline was 0.5-20μg/mL(r=0.9998,n=6).The limit of detection was 1.4 ng,and the limit of quantification was 5.0 ng.The RSDs of precision and stability test results were all lower than 2.0%.In the specificity test and the strong destructive test,oxytetracycline could be effectively separated from the degradation products of strong destruction and other eight known impurities.The detection rate of 2-acetyl-2 deamidothetracycline in Oxytetracycline Tablets was in the range of 1.62%-2.73%,and the total content of other impurities was in the range of 1.20%-1.84%.Conclusion The method is simple,has good specificity and high sensitivity,which can be used for the content determination of related substances in oxytetracycline API and Oxytetracycline Tablets.The limit for the relevant substance 2-acetyl-2-deamidothetracycline in the formulation of Oxytetracycline Tablets shall not exceed 3.5%of the main peak area of the reference solution,and the sum of peak areas of other impurities shall not exceed 2.0%of the main peak area of the reference solution.

关 键 词：土霉素 高效液相色谱法 有关物质 2-乙酰-2-去酰胺土霉素 

分 类 号：R917[医药卫生—药物分析学] R978.1[医药卫生—药学]