超高效液相色谱-串联质谱法快速测定化妆品中87种禁用原料

Rapid determination of 87 prohibited ingredients in cosmetics by ultra performance liquid chromatography-tandem mass spectrometry

建立了超高效液相色谱-串联质谱测定化妆品中87种禁用原料的方法,其适用于水基类、乳液膏霜类和油基类3种常见化妆品基质。实验对提取溶剂、提取时间及色谱、质谱条件进行了考察。水基类和乳液膏霜类化妆品用乙腈分散、50%乙腈水溶液超声提取,油基类化妆品用正己烷分散、70%乙腈水溶液涡旋提取;采用CORTECS C18色谱柱(150 mm×2.1 mm,2.7μm)进行分离,以乙腈和0.1%甲酸水溶液为流动相进行梯度洗脱,流速为0.3 mL/min,柱温为40℃,进样量为2μL。采用电喷雾电离(ESI)源,在多反应监测(MRM)模式下采集数据,利用基质匹配标准曲线进行定量分析。结果表明,87种禁用原料在各自的线性范围内具有良好的线性关系,相关系数(r)>0.99,检出限(LOD)为0.07~0.38μg/g,定量限(LOQ)为0.21~1.15μg/g。3种化妆品基质中87种禁用原料在低、中、高3个加标水平下的加标回收率为81.7%~115.4%,相对标准偏差(RSD,n=6)为0.4%~9.9%。采用该方法对349批化妆品样品进行测定,共发现8批阳性样品,检出成分分别为甲氧苄啶、特比萘芬、萘甲唑啉、7-甲氧基香豆素和7-甲基香豆素。本方法前处理过程简便,定性、定量可靠,可用于测定化妆品中87种禁用原料,能够为化妆品的快速风险筛查和国家标准的制修订提供技术支撑。

The methods of detecting numerous prohibited components are not included in the Technical Specifications for Cosmetic Safety(2015 Edition).Recently,owing to its high speed,sensitivity,and anti-interference properties,ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)became the preferred method of detecting banned substances in cosmetics.In this study,a UPLC-MS/MS method was developed for use in determining 87 prohibited ingredients in cosmetics,including 33 sex hormones,20 anti-infective drugs,15 antihistamines,7 coumarins,4 sedative-hypnotic drugs,4 antipyretic and analgesic drugs,2 allergenic fragrances,and 2 drugs with vasoconstriction effects.The main factors affecting the response,recovery,and sensitivity of the method,such as the type of extraction solvent,extraction time,ratio of the mobile phases,and MS conditions,were optimized during sample pretreatment and instrumental analysis.Accordingly,approximately 0.2 g of the toner or cream sample was dispersed in 2 mL acetonitrile in a 10 mL colorimetric tube.After diluting to 10 mL with 50%acetonitrile aqueous solution,the sample was ultrasonically extracted for 20 min and centrifuged,and the mixture was then filtered through a 0.22μm membrane.Approximately 0.2 g of the oil sample was dispersed in 2 mL n-hexane in a 15 mL polypropylene centrifuge tube and extracted twice with 3 mL 70% acetonitrile aqueous solution.The extracts were transferred into a 10 mL colorimetric tube and diluted to 10 mL with 50% acetonitrile aqueous solution,and the mixture was then filtered through a 0.22μm membrane.The samples were separated using a CORTECS C 18 column(150 mm×2.1 mm,2.7μm),employing a gradient elution program with acetonitrile and 0.1% formic acid aqueous solution as the mobile phases.The flow rate,column temperature,and injection volume were respectively set at 0.3 mL/min,40℃,and 2μL.The 87 compounds were monitored in multiple reaction monitoring(MRM)mode with electrospray ionization(ESI)under positive and negative conditions.Matrix-matched external standard calibration was used for quantification,and the analysis was completed within 33 min.The prohibited compounds exhibited good linear relationships,with r values of>0.99,and the limits of detection(LODs)and quantification(LOQs)for the 87 compounds were 0.07-0.38 and 0.21-1.15μg/g,respectively.Three types of cosmetic matrices were selected to verify the recovery and precision of the method at LOQ,2 LOQ,and 10 LOQ levels.The average recoveries of the 87 prohibited compounds were in the range of 81.7%-115.4%,and the relative standard deviations(RSDs,n=6)were 0.4%-9.9%.The reliability of the developed method was demonstrated by applying it to 349 commercial cosmetics obtained from the market,and 8 positive samples were identified.The positive components included trimethoprim,terbinafine,naphazoline,7-methoxycoumarin,and 7-methylcoumarin.The established method displays the advantages of simple operation and rapidness and a high sensitivity and good recovery.And,this method provides technical support for rapid risk screening and the revision of national standards for cosmetics.