中心切割二维液相色谱-质谱法定性鉴别盐酸特比萘芬乳膏中的未知杂质及其校正因子的测定

Qualitative Analysis of an Unknown Impurity in Terbinafine Hydrochloride Cream by Heart-cutting 2D-LC-MS and Determination of Its Correction Factor

关键词:  盐酸特比萘芬  中心切割  杂质  APCI  校正因子

Key words:  terbinafine hydrochloride  heart-cutting  impurity  APCI  correction factor

摘要:

目的 采用中心切割在线二维液相色谱四级杆飞行时间质谱法定性鉴别盐酸特比萘芬乳膏中相对保留时间（relative retention time，RRT）0.4的杂质并对其校正因子进行测定。方法 一维液相色谱采用Diamonsil C₁₈色谱柱，以三乙胺缓冲液（0.2%三乙胺溶液，用冰醋酸调节pH值至7.5）-甲醇-乙腈（30∶42∶28）为流动相A，以三乙胺缓冲液-甲醇-乙腈（5∶57∶38）为流动相B，梯度洗脱，流速为0.8 mL·min^-1，检测波长为280 nm；二维液相色谱采用Tsk gel ODS 100 V色谱柱，以0.1%甲酸水溶液-甲醇（20∶80）为流动相；质谱采用大气压化学电离离子源（APCI），正离子模式检测。根据一维液相色谱条件，对该杂质的校正因子进行测定。结果 根据质谱定性结果并结合对照品比对，确认盐酸特比萘芬乳膏中RRT 0.4的杂质为工艺杂质，可为后续精制工艺的改进提供依据。该杂质相对于特比萘芬的校正因子为0.21，提示定量时选择加校正因子的主成分自身对照法较为合理。结论 研究建立的中心切割在线二维液相色谱质谱法可用于盐酸特比萘芬乳膏中弱极性杂质的在线定性鉴别，为特比萘芬中其他弱极性杂质的鉴定提供了研究思路，也为复杂基质样品中微量杂质的分析鉴别提供了思路，可以为更好地监测盐酸特比萘芬乳膏的质量提供技术借鉴。

Abstract:

OBJECTIVE To establish a method of heart-cutting two-dimensional liquid chromatography coupled with Quadrupole time-of flight mass spectrometry(2D-LC-Q/TOF) to identify an impurity with relative retention time(RRT) of 0.4 in terbinafine hydrochloride cream and determine its correction factor. METHODS One-dimensional chromatographic conditions were as listed. A Diamonsil C₁₈ was used, and 0.2% triethylamine buffer(adjust pH to 7.5 with glacial acetic acid)-methanol-acetonitrile (30:42:28) as the mobile phase A, and triethylamine buffer-methanol-acetonitrile(5:57:38) as the mobile phase B, in gradient mode, at a flow rate of 0.8 mL·min^-1. The detection wavelength was set at 280 nm. Two-dimensional chromatographic conditions were as listed. A Tsk gel ODS 100 V was used, with 0.1% formic acid solution-methanol(20:80) as the mobile phase, APCI source was used at positive ion detection mode. The correction factor of the detected impurity was determined in the one-dimensional chromatographic system. RESULTS According to the qualitative results of mass spectrometry and the comparison of reference substance, the RRT 0.4 impurity in terbinafine hydrochloride cream was confirmed as the process impurity, which could provide the basis for the improvement of subsequent refining process. The correction factor of the impurity was 0.21 compared with terbinafine, which suggested that the method of self-control of main component with correction factor was more reasonable. CONCLUSION The 2D-LC-MS method can be used to identify weak polarity impurities in terbinafine hydrochloride cream, which provides ideas for the research of other weak polarity impurities in terbinafine hydrochloride cream and impurities of the similar preparations with complex matrix, and it is also an effect technology to control the quality of terbinafine hydrochloride cream.