柱后衍生化-HPLC法测定硫酸卡那霉素注射液及滴眼液的含量
|
摘要
目的新建硫酸卡那霉素注射液及滴眼液含量的高效液相色谱-柱后衍生化-荧光检测方法。方法色谱柱为ZorbaxEclipsePlusC18(4.6mm×100mm,3.5μm);流动相为pH3.4的缓冲液(取庚烷磺酸钠一水合物4.35g和无水硫酸钠16g,加水溶解并稀释至成1000mL,用冰醋酸调节pH值至3.4±0.1)-甲醇(74:26);柱温为35℃;流速为1mL/min。柱后衍生化试液为邻苯二甲醛溶液;流速为0.3mL/min;衍生化反应温度为45℃;荧光激发波长为340nm,发射波长为455nm。结果卡那霉素A在0.0973～583.8μg/mL浓度范围内线性关系良好(r=0.9999),硫酸卡那霉素注射液和滴眼液的平均回收率分别为99.6%和99.3%,RSD为0.63%和0.66%(n=9)。结论本方法检测灵敏度高,重复性好,简便快速,结果准确可靠,可作为硫酸卡那霉素注射液及滴眼液的含量测定方法。
Objective To establish a method of HPLC with post-column derivatization for the determination of the content in kanamycin sulfate injection and kanamycin sulfate eye drops. Methods A Zorbax Eclipse Plus C18 column(4.6 mm×100 mm, 3.5μm)was used, the mobile phase was buffer(containing 1-heptanesulfonate sodium salt and sodium sulfate)-methyl alcohol(74:26), the column temperature was 35℃, and the flow rate was 1 mL/min. The OPA solution was used for the post-column derivatization at a flow rate of 0.3 mL/min. The temperature of the postcolumn reaction was 45℃. The fluorescence was detected by an excitation and emission wavelength of 340 and 455 nm respectively. Results The calibration curve was linear in the range of 0.0973～583.8μg/mL(r=0.9999). The average recovery rates of kanamycin sulfate injection and kanamycin sulfate eye drops were 99.6% and 99.3%, and RSD values were 0.63% and 0.66%(n=9). Conclusion The method is rapid and versatile, and ensures excellent results in terms of sensitivity and reproducibility. It enables the content determination of kanamycin sulfate injection and kanamycin sulfate eye drops.
Objective To establish a method of HPLC with post-column derivatization for the determination of the content in kanamycin sulfate injection and kanamycin sulfate eye drops. Methods A Zorbax Eclipse Plus C18 column(4.6 mm×100 mm, 3.5μm)was used, the mobile phase was buffer(containing 1-heptanesulfonate sodium salt and sodium sulfate)-methyl alcohol(74:26), the column temperature was 35℃, and the flow rate was 1 mL/min. The OPA solution was used for the post-column derivatization at a flow rate of 0.3 mL/min. The temperature of the postcolumn reaction was 45℃. The fluorescence was detected by an excitation and emission wavelength of 340 and 455 nm respectively. Results The calibration curve was linear in the range of 0.0973～583.8μg/mL(r=0.9999). The average recovery rates of kanamycin sulfate injection and kanamycin sulfate eye drops were 99.6% and 99.3%, and RSD values were 0.63% and 0.66%(n=9). Conclusion The method is rapid and versatile, and ensures excellent results in terms of sensitivity and reproducibility. It enables the content determination of kanamycin sulfate injection and kanamycin sulfate eye drops.
引文
[1]李忠琴.卡那霉素A、B组分分离纯化新技术研究[D].福州:福州大学,2002.
[2]Umezawa H,Ueda M,Maeda K,et al.Production and isolation of a new antibiotic:Kanamycin[J].J Antibiot,1957,10(5):181-188.
[3]王金凤,杨化新,朱俐,等.HPLC-NQAD和HPLC-ELSD法测定硫酸卡那霉素注射剂的含量[J].药物分析杂志,2014,34(4):644-648.
[4]国家药典委员会.中华人民共和国药典(2015年版二部)[S].北京:中国医药科技出版社,2015:1316-1317.
[5]美国药典委员会.美国药典(第40版)[S].国家处方集,2017:4742-4744.
[6]赵敬丹,王曦,刘浩,等.柱后衍生化-HPLC法测定硫酸异帕米星注射液的含量和有关物质[J].中国抗生素杂志,2017,42(5):363-368.
[7]何丹,杨林.柱后衍生化高效液相色谱法测定妥布霉素滴眼液的含量[J].中国抗生素杂志,2010,35(10):771-773.
[8]于华生,陈东鸿,马赛骏.HPLC柱前衍生法测定妥布霉素[J].海峡药学,2001,13(4):28-30.
[9]薛晶,崔学文,胡昌勤.HPLC法测定人用皮卡狂犬病疫苗和皮卡佐剂中硫酸卡那霉素的含量[J].药物分析杂志,2011,31(10):1911-1913.
[2]Umezawa H,Ueda M,Maeda K,et al.Production and isolation of a new antibiotic:Kanamycin[J].J Antibiot,1957,10(5):181-188.
[3]王金凤,杨化新,朱俐,等.HPLC-NQAD和HPLC-ELSD法测定硫酸卡那霉素注射剂的含量[J].药物分析杂志,2014,34(4):644-648.
[4]国家药典委员会.中华人民共和国药典(2015年版二部)[S].北京:中国医药科技出版社,2015:1316-1317.
[5]美国药典委员会.美国药典(第40版)[S].国家处方集,2017:4742-4744.
[6]赵敬丹,王曦,刘浩,等.柱后衍生化-HPLC法测定硫酸异帕米星注射液的含量和有关物质[J].中国抗生素杂志,2017,42(5):363-368.
[7]何丹,杨林.柱后衍生化高效液相色谱法测定妥布霉素滴眼液的含量[J].中国抗生素杂志,2010,35(10):771-773.
[8]于华生,陈东鸿,马赛骏.HPLC柱前衍生法测定妥布霉素[J].海峡药学,2001,13(4):28-30.
[9]薛晶,崔学文,胡昌勤.HPLC法测定人用皮卡狂犬病疫苗和皮卡佐剂中硫酸卡那霉素的含量[J].药物分析杂志,2011,31(10):1911-1913.