臺灣博碩士論文加值系統

為了預防化粧品在長時間的使用過程中受到微生物汙染(例如：黴菌和酵母菌)，保障產品安全性，添加適量防腐劑是有其必要性。甲醛和5-bromo-5-nitro-1,3-dioxane被用在清潔類(或洗去型)化粧品中當作抗菌和防腐作用，特別是在洗面乳、洗髮精和沐浴乳中。甲醛於2006年被世界衛生組織WHO的國際癌症研究中心 ( International Agency for Research on Cancer)分類為第一類人類致癌物，5-bromo-5- nitro-1,3-dioxane (BND)則是一種含溴的抗菌化合物，國際上化粧品法規對其有不同的限量規定。因此，同時檢測化粧品中甲醛及5-bromo-5-nitro-1,3-dioxane的含量是有其必要性。本研究超高液相層析法可同時運用在不同清潔類化粧品中甲醛及5-bromo-5-nitro-1,3-dioxane的檢測，本方法僅將檢體經過簡單萃取並與乙醯丙酮衍生，不須經淨化或濃縮步驟便可以分析，檢驗方法確效以線性、準確度、精密度、定量極限及檢測極限來確保方法的性能，在最適化條件下，甲醛濃度在0.1到20 ppm之間，5-bromo-5-nitro-1,3-dioxane濃度在2到200 ppm之間，吸收光譜強度與濃度均呈線性關係，判定係數 (r2) 分別為1.0及0.9996，全部回收率均大於82 %，精密度之變異係數均小於8.2%，定量極限分別為0.1 ppm及2 ppm，檢測極限分別為0.02 ppm及0.8 ppm。本研究開發之方法具有寬的線性範圍，高選擇性和靈敏度，定量極限分別遠低於國際化粧品的規範，因此，本分析方法適用於市售清潔類化粧品中5-Bromo-5-nitro- 1,3-dioxane及甲醛的常規品質監測。

The addition of preservatives is absolutely essential to ensure that cosmetics are safe to use for a long time, since these compounds protect formulations from contamination by microorganisms such as bacteria and yeasts. 5-bromo-5-nitro-1,3-dioxane and formaldehyde are used in rinse-off cosmetics, particularly facial cleanser, shampoos and body wash, as antibacterial agent and preservatives. In 2006, International Agency for Research on Cancer (IARC) classified formaldehyde as carcinogen to humans. 5-Bromo - 5 - nitro-1 , 3 - dioxane is a bromine -containing preservative which also subjected to several restrictions according to the international cosmetic regulation. Thus, as a part of a quality control procedure, analytical methods for the determination of two compounds in different types of cosmetics are required. In this paper that ultra performance liquid chromatography has been simultaneously applied for the determination of formaldehyde and 5-bromo-5-nitro-1,3-dioxane in a variety of rinse off cosmetic products. After simple extraction, and derived with acetylacetone, it was analyzed without any further cleanup or concentration step. Linearity, accuracy, precision, limit of quantitation, and limit of detection were evaluated to assess the performance of the proposed method. Under optimal conditions, the spectrophotometry intensity increased linearly with the concentration of formaldehyde ranging from 0.1 to 20 ppm with a correlation coefficient of 1.0, and 5-bromo-5-nitro-1,3-dioxane ranging from 2 to 200 ppm with a correlation coefficient of 0.9996. The recovery studies on rinse-off cosmetics gave satisfactory values ( > 82 % for two analytes), and coefficient of variance of precision were all below 8.2 %. The quantitation limit of formaldehyde was 0.1 ppm, and 5-bromo-5-nitro-1,3-dioxane was 2 ppm. The detection limit of formaldehyde was 0.02 ppm, and 5-bromo-5-nitro-1,3-dioxane was 0.8 ppm. The proposed method has a wide linear range, high selectivity and sensitivity. The quantification limits were far below those set by the international cosmetic regulations, making this analytical method suitable for routine control.

目錄
中文摘要 I
Abstract V
目錄 VII
表目錄 XIV
縮寫對照表 XV
第一章 緒論 1
1-1 研究背景 1
1-1-1 甲醛 2
1-1-2 5-Bromo-5-nitro-1,3-dioxane 4
1-2 研究目的 10
第二章 法規與文獻探討 11
2-1 甲醛及供體的規範： 11
2-1-1 國際規範 11
2-1-2 國內規範 12
2-2 文獻探討 14
2-2-1 甲醛之檢測方法 14
2-2-2 5-Bromo-5-nitro-1,3-dioxane之檢測方法 19
第三章 材料與方法 22
3-1 材料 22
3-1-1 標準品 22
3-1-2 試藥 23
3-1-3 市售清潔類化粧品 24
3-1-4 器材 24
3-1-5 儀器設備 25
3-2 方法 26
3- 2-1 BND之UPLC層析條件 27
3- 2-2 BND之分析濃度範圍評估 29
3- 2-3 溫度對BND穩定性的影響 30
3- 2-4 pH值對BND穩定性的影響 31
3- 2-5 溶劑對清潔類化粧品中BND萃取率評估 32
3- 2-6 選擇衍生條件的最適化 33
3- 2-7 評估甲醛衍生需要的溫度與時間 35
3- 2-8 評估DDL於室溫下的穩定度 35
3- 2-9 溫度及衍生對BND及甲醛萃取的影響 36
第四章 結果與討論 38
4-1 BND之UPLC層析條件及分析濃度範圍評估 38
4-1-1 BND之UPLC層析條件 38
4-1-2 BND之分析濃度範圍評估 40
4-2 溫度及pH值對BND穩定性的影響 42
4-2-1 在溫度測試方面 42
4-2-2 在pH值測試方面 44
4-3 溶劑對清潔類化粧品中BND萃取率評估 47
4-3-1 卸粧水 47
4-3-2 清潔乳液 48
4-3-3 護髮霜 50
4-4 選擇衍生最適化的條件 52
4-4-1 最適化之衍生試劑組成 52
4-4-2 評估甲醛與乙醯丙酮試劑衍生的溫度與時間 59
4-4-3 DDL於室溫下的穩定度 60
4-5 溫度及衍生對BND及甲醛萃取的影響 62
4-5-1 溫度對BND及甲醛萃取的影響 62
4-5-2 衍生對BND及甲醛萃取的影響 62
4-6 分析方法確效 66
4-6-1 適用範圍(scope) 66
4-6-2 專一性(specificity)及線性(linearity) 67
4-6-3 準確度(accuracy) 71
4-6-4 精密度(precision) 74
4-6-5 定量極限(limit of quantification)及偵測極限(limit of detection) 77
第五章 結論 81
第六章 未來方向 84
參考文獻 85


圖目錄
圖1、 甲醛測試陽性反應結果的產品項目與件數 4
圖2、 Metabolism of 5-bromo-5-nitro-1,3-dioxane in rats and rabbits 8
圖3、 流動注射分光光度分析(FIA)衍生檢測系統的配置圖 16
圖4、 HPLC柱後衍生檢測系統的配置圖 17
圖5、 BND之化學結構圖 22
圖6、 甲醛之化學結構圖 23
圖7、 研究架構及流程圖 26
圖8、 使用移動相條件1分析BND標準溶液之層析圖 38
圖9、 使用移動相條件1分析添加BND卸粧水溶液之層析圖 39
圖10、 使用移動相條件2分析BND標準溶液之層析圖 39
圖11、 使用移動相條件1分析添加BND卸粧水溶液之層析圖 40
圖12、 BND標準溶液於40℃恆溫下時間與濃度的變化圖 43
圖13、 BND標準溶液於55℃恆溫下時間與濃度的變化圖 43
圖14、 室溫下濃度100 ppm BND於不同pH值之積分面積百分率 45
圖15、 室溫下濃度1000 ppm BND於不同pH值之積分面積百分率 45
圖16、 溶劑對卸粧水中BND萃取率之比較圖 48
圖17、 Methanol萃取卸粧水中BND之層析圖 48
圖18、 溶劑對清潔乳液中BND萃取率之比較圖 49
圖19、 Methanol萃取清潔乳液中BND之層析圖 49
圖20、 溶劑對護髮霜中BND萃取率之比較圖 50
圖21、 Methanol萃取護髮霜中BND之層析圖 50
圖22、 三種不同溶劑對於卸粧水、清潔乳液及護髮霜中BND之萃取率比較圖 51
圖23、 10 ppm甲醛溶液與不同冰醋酸含量之衍生試劑反應後的吸光值比較圖 53
圖24、 20 ppm甲醛溶液與不同冰醋酸含量之衍生試劑反應後的吸光值比較圖 54
圖25、 10 ppm及20 ppm甲醛衍生後吸光值與冰醋酸濃度的關係圖 55
圖26、 10 ppm甲醛溶液與不同醋酸銨含量之衍生試劑反應後的吸光值比較圖 56
圖27、 20 ppm甲醛溶液與不同醋酸銨含量之衍生試劑反應後的吸光值比較圖 56
圖28、 10 ppm及20 ppm甲醛衍生後吸光值與醋酸銨濃度的關係圖 57
圖29、 10 ppm甲醛溶液與不同乙醯丙酮含量之衍生試劑反應後的吸光值比較圖 58
圖30、 20 ppm甲醛溶液與不同乙醯丙酮含量之衍生試劑反應後的吸光值比較圖 58
圖31、 10 ppm及20 ppm甲醛衍生後吸光值與乙醯丙酮濃度的關係圖 59
圖32、 不同水浴溫度下，衍生反應與時間的關係圖 60
圖33、 室溫下DDL含量百分率與時間關係圖 61
圖34、 經衍生之甲醛及BND標準曲線與線性方程式 63
圖35、 不同溫度下、有無衍生BND之萃取率比較圖 64
圖36、 不同溫度下甲醛萃取率比較圖 65
圖37、 甲醛及BND兩標準曲線及判定係數 69
圖38、 甲醛 (10ppm) 及BND (100 ppm) 標準溶液之層析圖及吸收光譜 69
圖39、 甲醛 (0.1 ppm) 及BND (2.0 ppm) 標準溶液之層析圖 70
圖40、 添加標準品之卸粧水溶液的層析圖 70
圖41、 對照空白卸粧水溶液之層析圖 70
圖42、 對照空白溶液之層析圖 71
圖43、 甲醛與BND在兩種類型檢體之回收率比較圖 73
圖44、 甲醛及BND定量極限之層析圖 78
圖45、 卸粧水中添加甲醛及BND標準溶液濃度分別為0.05 ppm及1 ppm 78
圖46、 卸粧水中添加甲醛及BND標準溶液濃度分別為0.04 ppm及0.8 ppm 79
圖47、 卸粧水中添加甲醛及BND標準溶液濃度分別為0.025 ppm及0.5 ppm 79
圖48、 卸粧水中添加甲醛及BND標準溶液濃度分別為0.02 ppm及0.4 ppm 79
圖49、 卸粧水中添加甲醛及BND標準溶液濃度分別為0.0125 ppm及0.25 ppm 80

表目錄
表1、 美國市售不同類型產品中添加甲醛及供體種類之件數與百分率 6
表2、 美國FDA資料庫之化粧品產品中BND使用和濃度 7
表3、 歐盟、中華民國及日本對甲醛及其供體的使用濃度與規範 13
表4、 移動相體積比例與對應時間(條件1) 28
表5、 移動相體積比例與對應時間(條件2) 29
表6、 修改後之移動相體積比例與對應時間 39
表7、 BND標準曲線之濃度與線性範圍 41
表8、 衍生對BND及甲醛萃取率評估 62
表9、 BND及甲醛之添加回收率 73
表10、 樣品所含代測物之濃度範圍與回收率規範 74
表11、 樣品所含代測物之濃度範圍與變異係數規範 74
表12、 BND及甲醛於清潔乳液之重複性試驗 76
表13、 BND及甲醛於卸粧水之中間精密度試驗 77
表14、 最佳化之衍生及檢驗條件 82
表15、 分析方法之確效項目及其結果 83

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