高效液相色谱法测定硫唑嘌呤片中的2个有关物质

HPLC Determination of Two Related Substances in Azathioprine Tablets

目的:建立反相高效液相色谱法测定硫唑嘌呤片中的2个有关物质,6-巯基嘌呤和5-氯-1-甲基-4-硝基咪唑。方法:采用 ODS C_(18)柱(250 mm×4.6 mm,5μm)。流动相:庚烷磺酸钠溶液(取庚烷磺酸钠1.336 g,加水850 mL 溶解)-甲醇(85:15),用1 mol·L^(-1)盐酸调 pH 至3.5。流速:1.0 mL·min^(-1)。检测波长:254 nm。结果:6-巯基嘌呤浓度在4.68×10^(-4)～0.014 mg·mL^(-1)范围内与峰面积呈良好线性关系,回归方程为Y=5400X-0.5302,r=0.9997,平均回收率为99.5%,最低检测限1 ng;5-氯-1-甲基-4-硝基咪唑浓度在5.51×10^(-4)～0.0165 mg·mL^(-1)范围内与峰面积呈良好线性关系,回归方程为 Y=6743X-1.396,r=0.9994,平均回收率为99.7%,最低检测限2.5 ng。供试品溶液和对照品溶液均在8 h 内稳定。结论:本法专属性强、准确、灵敏、简便易行。

Objective： An HPLC method was developed to determine 6 -mercaptopurine and 5 -chloro -1 - methyl - 4 - nitroimidazole, which were related substances of azathioprine tablets. Methods： An ODS C18 column （250mm×4.6mm,5μm）was used with the mobile phase of a mixture of sodium heptanesulfonate solution（ sodium heptanesulfonate 1. 336 g,dissolved in 850 mL water） -methanol（85： 15,adjust the solution with 1 mol·L^-1 hydrochloric acid to a pH of 3.5）at a flow rate of 1.0 mL·min^-1 ;The detection wavelength was at 254 nm. Results：For 6 - mercaptopurine, the calibration curve was linear in the range of 4. 68 × 10^- 4 - 0. 014 mg·mL^-1 with correlation coefficient 0. 9997 and the regression equation being Y = 5400X - 0. 5302, the average recovery （ n = 9 ） was 99. 5% , and the detection limit was 1 ng ; For 5 - chloro - 1 - methyl - 4 - nitroimidazole, the calibration curve was also linear in the range of 5. 51 × 10^-4 -0. 0165 mg·mL^-1 with correlation coefficient 0. 9994 and the regression equation being Y= 6743X - 1. 396, the average recovery（n = 9） was 99.7% , and the detection limit was 2. 5 ng. Sample solution and standard solution are all stable within 8 h. Conclusion：This method is specific, accurate, sensitive and simple.