梯度洗脱HPLC法测定盐酸氨溴索口服溶液中甜味剂含量

目的: 建立高效液相色谱法测定盐酸氨溴索口服溶液中4种甜味剂(安赛蜜、糖精钠、阿司帕坦、甜菊糖/甜菊素)的方法,并在2011年度国家药品质量评价性抽验工作中,按照所建方法对盐酸氨溴索口服溶液中的4种甜味剂进行测定和评价。 方法: 采用Agilent SB-C₁₈(4.6 mm×150 mm,5 μm)柱,流动相A为0.02 mol·L^-1磷酸二氢铵缓冲液(pH 4.4),流动相B为乙腈,流速1.0 mL·min^-1,线性梯度洗脱,检测波长为200 nm(糖精钠、阿司帕坦、甜菊糖/甜菊素)和226 nm(安赛蜜)。 结果: 安赛蜜、糖精钠、阿司帕坦和甜菊糖/甜菊素分离完全,各辅料均无干扰,线性范围分别为0.00561~0.225 mg·mL^-1(r=1.0000)、0.00106~0.106 mg·mL^-1(r=1.0000)、0.00522~0.209 mg·mL^-1(r=0.9999)和0.00517~0.296 mg·mL^-1(r=1.0000);检测限分别为0.4 ng,0.4 ng,1.6 ng和15.5 ng;定量限分别为1.3 ng,1.5 ng,5.2 ng和51.7 ng;平均加样回收率(n=6)分别为98.8%(RSD=0.5%),99.5%(RSD=0.3%),100.6%(RSD=0.2%),99.1%(RSD=1.1%)。 结论: 本法具有较高的选择性,结果稳定,通过测定甜味剂的含量可监控制剂生产过程中辅料投料是否与处方一致。

Objective: To develop a method for determination of sweeteners in ambroxol hydrochloride oral solution including potassium acesulfame,sodium saccharine,aspartame and stevioside/steviosin. Methods: The separation was conducted on an Agilent SB-C₁₈(4.6 mm×150 mm,5 μm)and coupled with DAD at 200 nm(sodium saccharine,aspartame and stevioside/steviosin)and 226 nm(potassium acesulfame).The mobile phase consisted of 0.02 mol·L^-1 ammonium dihydrogen phosphate(pH 4.4)(A)and acetonitrile(B)with gradient program.The flow rate was 1.0 mL·min^-1. Results: Good separation of 4 sweeteners and other excipients were achieved.The linear ranges of potassium acesulfame,sodium saccharinem,aspartame and stevioside were 0.00561-0.225 mg·mL^-1 (r=1.0000),0.00106-0.106 mg·mL^-1 (r=1.0000),0.00522-0.209 mg·mL^-1 (r=0.9999),and 0.00517-0.296 mg·mL^-1 (r=1.0000),respectively.The limits of detection were 1.3 ng,1.5 ng,5.2 ng,and 51.7 ng,respectively.The average recoveries (n=6) were 98.8% (RSD=0.5%),99.5 % (RSD=0.3%),100.6 % (RSD=0.2%),and 99.1% (RSD=1.1%),respectively. Conclusions: The method showed high selectivity,and sensitivity.The determination results can be applied for monitoring of the manufacturing process.