Resident determination of benzyltrimethylammonium hydroxide in bemiparin sodium by HPLC
Shao Fang1,3, Ren Fengzhi1,3, Liu Jianfen2,3, Wang Yue1,3, Han Yang1,3, Li Ning1,3
1. North China Pharmaceutical Group Corporation, New Drug Research and Development Corporation Ltd., Shijiazhuang 052165, China; 2. North China pharmaceutical Huakun Hebei Biotechnology Corporation Ltd. ,Shijiazhuang 050000, China;
3. Hebei Multi-component Biochemical Drug Preparation Technology Innovation Center, Shijiazhuang 050000, China
Abstract：An HPLC method for determination of Benzyltrimethylammonium hydroxide was established in Bemiparin sodium. The chromatographic conditions were determined as follows: the analysis is performed on Agilent ZORBAX Eclipse XDB-CN column（250×4.6 mm,5 μm）, the column temperature is set to 35 ℃ , 0.2 mol·L-1 sodium perchlorate solution(Adjusting pH vaule of 2.2 with 1 mol·L-1 perchloric acid)-acetonitrile（88∶12）is adopted as mobile phase, and the flow rate of 0.8 mL·min-1, the detection wavelength of 207 nm, and the sample volume of 50 μL are adopted. The results show that the concentration of benzyltrimethylammonium hydroxide bears a good linear relationship with peak area within the range of 0.01 to 1.0 μg·mL-1 (r=0.999 8, n=7); the RSD of the repeatability test results is 1.21% (n=6); the RSD of the intermediate precision test results is 1.35% (n=12); the RSD of the robustness test results is 1.20% (n=16); the stability of the solution is good within 48 hours with an RSD of 0.19% (n=13); the average recovery rate is 102.40%, with an RSD of 1.70% (n=9). This method is simple to operate, highly sensitive, and provides accurate and reliable results. It can be used for the determination of benzyltrimethylammonium hydroxide residues in Bemiparin sodium.
