盐酸地巴唑原料药和片剂中有关物质的测定及最大未知杂质结构的推测  

Determination of Related Substances in Dibasol Hydrochloride Raw Materials and Tablets and Structure Prediction of Maximum Unknown Impurity

作　　者：陈宇堃[1] 梁蔚阳[1] CHEN Yukun;LIANG Weiyang(Guangdong Institute for Drug Control/NMPA Key Laboratory for Quality Control of Blood Products/Guangdong Drug Administration Key Laboratory of Quality Control and Research of Blood Products,Guangzhou 510663,China)

机构地区：[1]广东省药品检验所/国家药品监督管理局血液制品质量控制重点实验室/广东省药品监督管理局血液制品质量控制研究重点实验室,广州510663

出　　处：《中国药房》2020年第6期696-702,共7页China Pharmacy

基　　金：广东省科技计划项目(No.2015A030402004);广东省医学科学技术研究基金项目(No.B2018130,No.B2018220);广东省食品药品监督管理局科技创新项目(No.2019ZDZ23,No.2018YDB02)。

摘　　要：目的:建立测定盐酸地巴唑原料药及片剂中有关物质的方法,并对其最大未知杂质的结构进行推测。方法:采用高效液相色谱法测定盐酸地巴唑原料药及片剂中的有关物质(邻苯二胺、苯乙酸),色谱柱为KromasilC18,流动相为水-甲醇-冰醋酸-三乙胺(45∶55∶0.5∶0.5,V/V/V/V),流速为1.0 mL/min,检测波长为220 nm,柱温为30℃,进样量为10μL。采用超高效液相色谱-飞行时间质谱法、氢谱、碳谱推测最大未知杂质的结构,色谱柱为Waters Acquity UPLC BEH C18,流动相为水-甲醇(45∶55,V/V),流速为0.2 mL/min,柱温为30℃,进样量为1μL,离子源为电喷雾离子源(ESI),扫描模式为负离子扫描模式,一级质谱扫描范围为m/z 100~800,毛细管电压为3000 V,离子源温度为100℃,去溶剂气体为氮气,去溶剂气体流速为600 L/h,锥孔流速为50 L/h。结果:邻苯二胺、苯乙酸、盐酸地巴唑检测质量浓度的线性范围为0.427~4.27μg/mL(r=0.9989)、0.403~4.03μg/mL(r=0.9989)、0.82~8.20μg/mL(r=0.9999);定量限分别为0.0427、0.1343、0.0887μg/mL,检测限分别为0.0214、0.0671、0.0443μg/mL;精密度、稳定性、重复性、耐用性试验的RSD均小于2%;邻苯二胺的加样回收率为98.31%~102.81%(RSD=1.60%,n=9)、苯乙酸为99.78%~102.23%(RSD=0.70%,n=9)。6批盐酸地巴唑原料药中均未检出邻苯二胺,苯乙酸含量为0~0.04%,最大未知杂质含量为0.05%~0.25%,未知杂质总量为0.05%~0.31%;77批盐酸地巴唑片剂中,邻苯二胺含量为0~0.11%,苯乙酸含量为0~0.03%,最大未知杂质含量为0.06%~0.51%,未知杂质总量为0.10%~0.62%。推测最大未知杂质为2-(羟苯基甲基)苯并咪唑(羟苄唑)。结论:所建方法快速、准确、专属性好,可用于测定盐酸地巴唑原料药及片剂中的有关物质。最大未知杂质可能为苯并咪唑类化合物。OBJECTIVE:To establish a method for the determination of related substances in dibasol hydrochloride raw materials and tablets, and to predict the maximum unknown impurity’s structure. METHODS: The related substances(o-phenylenediamine,phenylacetic acid)in dibasol hydrochloride raw materials and tablets were determined by HPLC. The determination was performed on Kromasil C18 column with mobile phase consisted of mobile phase-methanol-glacial acetic acid-triethylamine(45 ∶ 55 ∶ 0.5 ∶ 0.5,V/V/V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 220 nm,and column temperature was 30 ℃. Sample size was 10 μL. UPLC-TOF-MS,1H-NMR and 13C-NMR were used for structure prediction.The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of water-methanol(45 ∶ 55,V/V) at the flow rate of 0.2 mL/min. The column temperature was 30 ℃,and sample size was 1 μL. The ion source was electrospray ion source. The scanning mode was negative ion scanning mode. The first-order mass spectrum scanning range was m/z100-800,the capillary voltage was 3 000 V,the source temperature was 100 ℃,the desolvent gas was nitrogen,and the solvent free gas flow rate was 600 L/h. The flow rate of the conical orifice was 50 L/h. RESULTS: The linear range of o-phenylenediamine,phenylacetic acid and dibasol hydrochlo-ride were 0.427-4.27 μ g/mL(r=0.998 9),0.403-4.03 μ g/mL(r=0.998 9)and 0.82-8.20 μg/mL(r=0.999 9),respec-tively. The limits of quantitation were 0.042 7,0.134 3,0.088 7 μg/mL. The limits of detection were 0.021 4,0.067 1,0.044 3 μ g/mL.RSDs of precision,stability,reproducibility and durability tests were all less than 2%. The average recoveries were 98.31%-102.81% for o-phenylenediamine(RSD=1.60%,n=9),and99.78%-102.23% for phenylacetic acid(RSD=0.70%,n=9).No o-phenylenediamine was detected in 6 batches of dibazol hydrochloride raw materials;the contents of phenylacetic acid were 0-0.04%;the contents of maximum unknown impurity were 0.05%-0.25%;total contents of un

关 键 词：盐酸地巴唑原料药 盐酸地巴唑片剂 高效液相色谱法 有关物质 超高效液相色谱-飞行时间质谱法 杂质 结构鉴定 

分 类 号：R927.1[医药卫生—药学]