复方他扎罗汀丙酸氯倍他索软膏有关物质的色谱-质谱结构鉴定

目的：采用色谱-质谱联用技术鉴定新型外用制剂复方他扎罗汀丙酸氯倍他索软膏中的有关物质，为其质量研究提供参考依据。方法：采用Phecda C₁₈色谱柱（4.6 mm×250 mm，5 μm），以0.1%乙酸铵/0.2%甲酸水溶液-甲醇-乙腈（70：21：9）为流动相A，甲醇-乙腈（70：30）为流动相B，进行线性梯度洗脱，检测波长为240 nm和325 nm，分别对丙酸氯倍他索和他扎罗汀的有关物质进行检测；采用电喷雾正离子化-飞行时间质谱法（ESI⁺-TOF/MS）测定各有关物质的准确质量和元素组成，三重四极杆串联质谱测定子离子特征，解析鉴定有关物质的结构。结果：在所建立的条件下，复方他扎罗汀丙酸氯倍他索软膏中的主成分他扎罗汀、丙酸氯倍他索及有关物质分离良好，检测并鉴定出12个有关物质，其中7个为他扎罗汀有关物质，5个为丙酸氯倍他索有关物质；鉴定出有关物质2、5、6、8、9、11和12分别为他扎罗汀亚砜、他扎罗汀酸、他扎罗汀酸甲酯、倍他米松17-丙酸酯、氯倍他索、倍他米松21-丙酸酯和17α-螺环化合物，均为已知有关物质，而未知有关物质1、3、4、7和10为本研究首次鉴定。结论：建立的色谱-质谱联用方法能有效地分离鉴定复方他扎罗汀丙酸氯倍他索软膏中的有关物质，鉴定结果为其质量控制提供了参考。

Objective: To develop LC-MS methods for identification of the related substances of compound tazarotene and clobetasol propionate ointment,and provide a reference for its quality control. Methods: The separation was performed on a Phecda C₁₈ column (4.6 mm×250 mm,5 μm) by linear gradient elution using 0.1% ammonium acetate/0.2% formic acid buffer solution-methanol-acetonitrile (70:21:9) as mobile phase A and methanol-acetonitrile (70:30) as mobile phase B. The detection wavelength for clobetasol propionate and tazarotene were 240 nm and 325 nm,respectively. The exact mass and elemental composition of the parent ions of each related substance were determined by high resolution TOF/MS operated in electrospray positive mode,triple quadrupole tandem mass was employed for the product mass spectra determination,and the structures of the related substances were analyzed and identified. Results: The principal components of compound tazarotene and clobetasol propionate ointment,tazarotene,clobetasol propionate and its related substances were adequately separated under the established conditions. Twelve related substances were detected and identified,seven of them were related to tazarotene,the others were the related substances of clobetasol propionate. The related substances 2,5,6,8,9,11 and 12 were identified as tazarotene sulfoxide,tazarotenic acid,tazarotenic acid methylate,betamethasone 17-propionate,clobetasol,betamethasone 21-propionate and 17α-spiro compound,respectively,all of which are known related substances,and the unknown related substances 1,3,4,7 and 10 were identified for the first time in this paper. Conclusion: The developed LC-MS methods are effective for the separation and identification of related substances of compound tazarotene and clobetasol propionate ointment,the identification results provide references for its quality control.

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