化粧品中染髮劑成分檢驗方法之建立   全文下載

Establishment of the Analytical Method for Hair Dyes in Cosmetics

本研究以液相層析串聯質譜法（Liquid Chromatography/Tandem Mass Spectrometry, LC-MS/MS）建立化粧品中5項染髮劑成分之定量檢驗方法，利用電灑游離法（Electrospray Ionization, ESI）搭配多重反應偵測模式（Multiple Reaction Monitoring, MRM），並以Cortecs UPLC C18+（1.6μm, 2.1×100 mm）作為層析管柱，檢測化粧品中2-Chloro-6-ethylamino-4-nitrophenol、HC violet No.2、Hydroxybenzomorpholine、4-Nitrophenyl aminoethylurea及6-Methoxy-2-methylamino-3-aminopyridine hydrochloride等5項成分。研究中以含0.1%抗壞血酸之50%甲醇溶液萃取檢體，再經LC-MS/MS分析，以pH 3.5之10 mM甲酸銨水溶液及甲醇：乙腈（1：1, v/v）溶液作為移動相，進行梯度沖提，層析時間為15分鐘，即可獲得良好分析結果。另以市售染髮產品作為基質進行方法確效，結果顯示基質添加標準品均無干擾，具良好專一性。HC violet No.2、Hydroxybenzomorpholine及4-Nitrophenyl aminoethylurea之定量極限為2μg/g，2-Chloro-6-ethylamino-4-nitrophenol及6-Methoxy-2-methylamino-3-aminopyridine hydrochloride之定量極限則為4μg/g，前述5項成分之檢量線相關係數（Correlation coefficient, r）均可達0.9968以上，平均回收率介於82.2-102.7%，變異係數均小於13.5%，顯示方法具有良好之準確度及精密度。以14件市售染髮產品進行評估，結果顯示本方法可適用相關產品檢驗。未來將公開本研究之檢驗方法，供各界參考使用。

This study develops a quantitative analytical method for the detection of five hair dye ingredients in cosmetic products using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method utilizes electrospray ionization (ESI) coupled with multiple reaction monitoring (MRM), employing a Cortecs UPLC C18+ column (1.6μm, 2.1 x 100 mm) for chromatographic separation. The five target compounds include 2-chloro-6-ethylamino-4-nitrophenol, HC violet No.2, hydroxybenzomorpholine, 4-nitrophenyl aminoethylurea, and 6-methoxy-2-methylamino-3-aminopyridine hydrochloride could be analyzed by the method. Samples were extracted with 50% methanol solution containing 0.1% ascorbic acid prior to LC-MS/MS analysis. Gradient elution was performed using 10 mM ammonium formate aqueous solution at pH 3.5 and methanol: acetonitrile (1:1, v/v) solution, with a chromatographic run time of 15 minutes, yielding good analytical results. The method was validated using commercially available hair dye products as matrices, and the results showed no interference from added standards, demonstrating good specificity. The limits of quantification (LOQs) for HC violet No. 2, hydroxybenzomorpholine, and 4-nitrophenyl aminoethylurea were 2μg/g, while the LOQs for 2-chloro-6-ethylamino-4-nitrophenol and 6-methoxy-2-methylamino-3-aminopyridine hydrochloride were 4μg/g. The correlation coefficients (r) for the calibration curves of these five compounds exceeded 0.9968. Average recovery rates ranged from 82.2% to 102.7%, and the coefficients of variation were all below 13.5%, indicating good accuracy and precision. A total of 14 hair dye products were evaluated, and the results demonstrated that the established method is applicable for the analysis of related products. In the future, this established testing method will be announced for reference and used by relevant parties.