三氟乙醯丁酯衍生法於化妝品中胺基酸成分之GC-MS鑑定應用

Trifluoroactyl-Butyl Ester Derivatization for the GC-MS Identification of Amino Acids in Cosmetics

朱靜慧(Ching-Hui Chu) ； 徐照程(Jaw-Cherng Hsu)

《弘光學報》 55期 (2009/04) Pp. 151-163

https://doi.org/10.6615/HAR.200904.55.12

摘要

本研究採用正丁醇與氯化乙醯試劑經110℃，30分鐘的羧基(carboxylate group)烷化反應，與三氟醋酸酐(TFAB)試劑經140℃，10分鐘的胺基(amino group)醯化反應衍生微量胺基酸混合標準品，使其成為安定易揮發之TFAB衍生物，注入裝置SGE Forte ID-BPX5管柱之GC-MS系統，在起始溫度50℃，100℃/min升至250℃並持續10分鐘的條件下，23分鐘內能有效鑑定十六種胺基酸TFAB衍生物。具非極性側鏈與具有羧基與胺基側鏈的胺基酸擁有較高的反應產率，推測非極性溶劑環境有利於前者的反應，後者則歸因於羧基或胺基側鏈提供了更多的烷化或醯化反應位置。相反的，其他極性側鏈之胺基酸則產率偏低，但本系統仍能對其靈敏鑑別。大部分胺基酸分析再現性的相對標準偏差值皆可低於20%；其中Methionine、Glutamic acid及Aspartic acid的相對標準偏差值分別為8.8、11.0、11.4%，是較佳的前三名；在定量實驗中，部份胺基酸在50~200ppm範圍內具有良好的線性關係。本方法進一步應用至市售複合胺基酸保濕原料Prodew的分析，鑑定出alanine、glycine、threonine等七種胺基酸成分的存在；而Lipacide檢出含有alanine、glycine、valine等多達十種胺基酸成份，證明本研究對化妝品中胺基酸組成鑑定的能力與價值。

關鍵字

烷化 ； 醯化 ； 胺基酸衍生 ； GC-MS ； 化妝品

並列摘要

A selective and sensitive method for determination of amino acids in cosmetics by GC-MS has been developed. The amino acids in samples were directly converted into their trifluoroacetyl-butyl ester (TFAB) derivatives by consecutive alkylation of its carboxylate groups and acylation of its amino groups, and measured by GC-MS using a SGE Forte ID-BPX5 column. Using this method, the TFAB-amino acids derivatives provided excellent response and were quantitatively and reproducibly resolved within 23 min. Higher yield were observed in the derivation of amino acids with non-polar or carboxylate or amino side chain. The inter-assay R.S.D. of most amino acids were lower than 20% (n=3). The calibration curves for some amino acids in the range 50-200 ppm were linear and sufficiently reproducible for quantitative analysis. This method was successfully applied to commercial cosmetic raw materials without prior cleanup; there were no evidence of interference from coexisting substances from such a complex sample matrix.

並列關鍵字

Amino acid ； cosmetics ； GC-MS ； Derivatization GC ； Alkylation ； Acylation

參考文獻

K. Katayama, J. A. Borunda, R. Rachow, 1993. A pentapeptide from type I procollagen promotes extracellular matrix production. J. Biol Chem, 268, 9941-9944.

L. M. Gutierrez, S. Viniegra, J. Rueda, 1997. A peptide that mimics the C-terminal sequence of SNAP-25 inhibits secretory vesicle docking in chromaffin cells. J. Biol. Chme, 272, 2634-2639.

A. Aitken, M. J. Geisow, J. B. C. Findlay, C. Holmes, A. Yanuaod, 1989. Protein Sequencing A Practical Approach, 43-51.

T. Bergman, M. Csrlquist, H. Jornvall, 1986, in Advanced Methods in Protein Microsequencer Analysis, 45-55.

I.G. Casella, M. Contursi, 2003, Isocratic ion chromatographic determination of underivatized amino acids by electrochemical detection. Analytica Chimica Acta 478, 179-189.