论文部分内容阅读
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 establish a RP-HPLC method for the determination of related substances of β-stavudine and a solution method for preparing system suitability test by hydrolytic destruction. Methods: An Agilent 1100 High Performance Liquid Chromatography (HPLC) was performed on a SUPELCOSIL LC-18-DB (4.6 mm × 250 mm, 5 μm) column with 0.01 mol·L -1 ammonium acetate solution-acetonitrile mol·L -1 ammonium acetate solution - acetonitrile (75:25) as mobile phase, gradient elution, detection wavelength 254 nm, column temperature 25 ℃. Results: β-stavudine and four known impurities and other unknown impurities were able to be effectively separated; the separation of α-stavudine and β-stavudine produced by hydrolysis and destruction reached 2.8; β The linear ranges of stavudine, thymine, β-thymidine and 5-O’-benzoyl-stavudine were 0.51 ~ 26μg · m L -1 (r = 1.000), 0.13 ~ 27μg · m L-1 (r = 1.000), 0.50-25 μg · m L -1 (r = 1.000) and 1.7-6.3 μg · m L -1 (r = 1.000). Thymine, (RSD = 1.5%), 100.6% (RSD = 0.9%), 101.9% (RSD = 2.1%) and the average recoveries (n = ); the minimum detectable amount of β-stavudine and the three known impurities was below 2.5 ng; the reproducibility of the system suitability solution prepared by hydrolytic destruction was good; the test solution was stable for 30 h at 4 ° C Within the basic stability. Conclusion: The method is sensitive, accurate, reliable and specific. It can be used for the determination of related substances of β-stavudine.