RP-HPLC法测定β-司他夫定的有关物质  

RP-HPLC determination of related substances in β-stavudine

作　　者：吴颖[1] 张小红[1] 耿志旺[1] 马玲 乐健[1] 杨永健[1] 

机构地区：[1]上海市食品药品检验所,上海201203 [2]上海迪赛诺生物医药有限公司,上海201203

出　　处：《药物分析杂志》2015年第1期103-108,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的：建立了β-司他夫定原料有关物质的RP-HPLC测定方法和水解破坏制备系统适用性试验溶液方法。方法：采用Agilent 1100型高效液相色谱仪,使用SUPELCOSIL LC-18-DB（4.6 mm×250 mm,5μm）色谱柱,以0.01 mol·L^-1醋酸铵溶液-乙腈（96.5∶3.5）和0.01 mol·L^-1醋酸铵溶液-乙腈（75∶25）为流动相,梯度洗脱,检测波长254 nm,柱温25℃。结果：β-司他夫定和4种已知杂质及其他未知杂质均能达到有效分离;经水解破坏产生的α-司他夫定与β-司他夫定的分离度均达2.8;β-司他夫定、胸腺嘧啶、β-胸苷与5-O＇-苯甲酰-司他夫定线性范围分别为0.51-26μg·m L^-1（r=1.000）、0.13-27μg·m L^-1（r=1.000）、0.50-25μg·m L^-1（r=1.000）、1.7-6.3μg·m L^-1（r=1.000）,已知杂质胸腺嘧啶、β-胸苷与5-O＇-苯甲酰-司他夫定的平均加样回收率（n=9）分别为102.8%（RSD=1.5%）、100.6%（RSD=0.9%）、101.9%（RSD=2.1%）;β-司他夫定与3种已知杂质的最小检出量均在2.5 ng以下;经水解破坏制备的系统适用性溶液的重复性良好;供试品溶液在4℃下的30 h内基本稳定。结论：本方法灵敏、准确、可靠,专属性强,可用于β-司他夫定原料的有关物质测定。Objective： To develop a method for RP-HPLC determination of related substances in β-stavudine and prepare the solution for system suitability test by sample hydrolysis. Methods： The chromatographic separation was achieved on a SUPELCOSIL LC-18-DB column（ 4. 6 mm × 250 mm,5 μm） with Agilent 1100 liquid chromatography system. The mobile phases consisted of 0. 01 mmol·L^-1ammonium acetate solution-acetonitrile（ 96. 5∶ 3. 5）and 0. 01 mmol·L^-1ammonium acetate solution-acetonitrile（ 75 ∶ 25） with gradient elution. The detection wavelength was 254 nm,and the column temperature was 25 ℃. Results： Good separation between β-stavudine and 4 identified impurities and other unknown impurities could be achieved,the resolution between β-stavudine and α-stavudine produced by hydrolysis was 2. 8. Good linear relationship was observed in the concentration range of0. 51-26 μg·m L^-1（ r = 1. 000） for β-stavudine,0. 13-27 μg·m L^-1（ r = 1. 000） for thymine,0. 50-25 μg·m L^-1（ r = 1. 000） for β-thymidine and 1. 7-6. 3 μg·m L^-1（ r = 1. 000） for 5-O＇-benzoyl-stavudine. The average recoveries of thymine,β-thymidine and 5-O＇-benzoyl-stavudine were 102. 8%（ RSD = 1. 5%）,100. 6%（ RSD = 0. 9%） and 101. 9%（ RSD = 2. 1%）,respectively. The limits of detection of β-stavudine and identified impurities were all below 2. 5 ng·m L^-1. The solution for system suitability test prepared by hydrolysis also showed good reproducibility. Sample solutions were stable at 4 ℃ in 30 hours. Conclusion： This method is sensitive,accurate,specific and reproducible,which can serve as a reliable analytical method for the determination of related substances in β-stavudine.

关 键 词：β-司他夫定 杂质检测 胸腺嘧啶 β-胸苷 5-O'-苯甲酰-司他夫定 α-司他夫定 水解破坏产物 有关物质 高效液相色谱 

分 类 号：R917[医药卫生—药物分析学]