Citation: Cai-Sheng Wu, Cai-Hong Wang, Jin-Lan Zhang, Dong-Mei Wang, Yuan-Feng Tong, Song Wu, Hai-Wei Huang, Bao-Ming Ning. Separation, determination of six impurities in methotrexate drug substance using ultra-performance liquid chromatography[J]. Chinese Chemical Letters, ;2014, 25(3): 447-450. doi: 10.1016/j.cclet.2013.11.048 shu

Separation, determination of six impurities in methotrexate drug substance using ultra-performance liquid chromatography

1.
a State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China;
2.
b Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China;
3.
c National Institutes for Food and Drug Control, Beijing 100050, China

Corresponding author: Jin-Lan Zhang,
Received Date: 30 August 2013
Available Online: 14 November 2013
Fund Project: We thank the Ministry of Public Health of the People's Republic of China (No. 200802038) (No. 200802038)

Methotrexate (MTX) is an antineoplastic therapeutic medicine as antimetabolite of folic acid. In this paper, a sensitive and rapid ultra-performance liquid chromatographic (UPLC) method was developed and validated for the separation and determination of impurities in MTX drug substances. The UPLC method was accomplished on an Agilent Zorbax Extend C-18 (50 mm×4.6 mm, 1.8 mm) with a gradient elution system composed of sodium dihydrogen phosphate in water (20 mmol/L, pH 3.0) and acetonitrile. The flow rate was 2.2 mL/min. The method was validated. The calibration curves displayed good linearity (r > 0.999) within the tested concentration ranges. The limit of detection (LOD) and limit of quantification (LOQ) of the six analytes were all less than 0.774 μg/mL and 1.03 mg/mL. The relative standard deviation (RSD) for intra-and inter-day precision of the six analytes was less than 9.8%, including at the LOQ. The average recovery ranged from 95.2% to 103% except at the LOQ, where recovery ranged from 82.7% to 117%. The validated method was successfully used to determine the relative abundance of six impurities in the MTX drug substances.
- Methotrexate,
- Ultra-performance,
- Liquid chromatography,
- Impurity
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沈阳化工大学材料科学与工程学院沈阳 110142