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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 Δ

陈宇堃 ，梁蔚阳（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）

中图分类号 R927.1 文献标志码 A 文章编号 1001-0408（2020）06-0696-07
DOI 10.6039/j.issn.1001-0408.2020.06.12

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10 μL。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1 μL，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3 000 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0.427～4.27 μg/mL（r＝0.998 9）、0.403～4.03 μg/mL（r＝0.998 9）、
0.82～8.20 μg/mL（r＝0.999 9）；定量限分别为0.042 7、0.134 3、0.088 7 μg/mL，检测限分别为0.021 4、0.067 1、0.044 3 μg/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。
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

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

ABSTRACT 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，H-NMR and C-NMR were used for structure prediction.
1
13
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 μ L. The ion source was
electrospray ion source. The scanning mode was negative ion scanning mode. The first-order mass spectrum scanning range was m/z
100-800，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 μg/mL（r＝0.998 9），0.403-4.03 μg/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.
Δ 基金项目：广东省科技计划项目（No.2015A030402004）；广东
RSDs of precision，stability，reproducibility and durability tests
省医学科学技术研究基金项目（No.B2018130，No.B2018220）；广东省
were all less than 2％. The average recoveries were 98.31％-
食品药品监督管理局科技创新项目（No.2019ZDZ23，No.2018YDB02）
102.81％ for o-phenylenediamine（RSD＝1.60％ ，n＝9），and
＊副主任药师。研究方向：生化药品和生物制品质量控制。电
话：020-81848209。E-mail：bettycyk@163.com 99.78％-102.23％ for phenylacetic acid（RSD＝0.70％，n＝9）.
# 通信作者：主任药师，硕士。研究方向：生化药品和生物制品评 No o-phenylenediamine was detected in 6 batches of dibazol
价与质量控制。电话：020-81848209。E-mail：wl-1023@163.com hydrochloride raw materials；the contents of phenylacetic acid

·696 · China Pharmacy 2020 Vol. 31 No. 6 中国药房 2020年第31卷第6期