以液相層析串聯式質譜技術檢測肉類製成品中的雜環胺化合物

Analysis of Heterocyclic Aromatic Amines in Meat Products by Liquid Chromatography - Tandem Mass Spectrometry

簡詩涵(Jian Shi-Han)

指導教授 ： 陳頌方

國立臺灣師範大學/理學院/化學系/碩士(2017年)

https://doi.org/10.6345/NTNU202202908

摘要

雜環胺化合物已經被證實可能在人體中會導致癌症發生，國際癌症研究機構(The International Agency for Research on Cancer, IARC)將MeIQ、MeIQx及PhIP歸類為2B類致癌物，IQ則歸類為2A類致癌物，2011年國家毒理學計畫(National Toxicology Program)指出MeIQ、MeIQx、IQ及PhIP也皆可被合理預期為誘發人類癌症的物質。在平常家庭烹煮肉類條件下，約150℃下烹煮如牛肉、豬肉及魚類等蛋白質含量豐富的肉類即會產生雜環胺化合物。為達最佳萃取效率，針對肉類樣品中的11種雜環胺化合物進行樣品前處理優化，開發並比較兩種樣品前處理法，第一種為液相萃取(Liquid-Liquid Extraction, LLE)法搭載固相萃取法(Solid-Phase Extraction, SPE)之樣品前處理法，第二種為較新穎的QuEChERS樣品前處理法，經過萃取後搭配高效能液相層析及串聯式質譜儀(HPLC-MS/MS)進行檢測。使用乙腈做為液相萃取溶劑，接著使用Oasis® MCX的管柱進行萃取，QuEChERS法首先使用乙腈做液相微萃取溶劑，C18EC、PSA及MgSO4做為分散式固相萃取吸附劑(dSPE)。11種雜環胺化合物在此兩種方法驗證下得到很好的精密度(RSD < 15.15%)、準確率(79.80-117.64%)、回收率(52.39-116.88%)、偵測極限(0.01-10 ppb)及線性回歸之相關係數(r > 0.993)。此外，QuEChERS樣品前處理法具有較好的線性動態範圍，因此具有更高的靈敏度。此兩種法也成功應用在檢測市面上肉類製成品(肉鬆及肉乾)中的雜環胺化合物。在六種樣品中可定量出三至四種的雜環胺化合物，定量出的濃度範圍為0.06-718.46ppb。

關鍵字

雜環胺化合物 ； 液相層析-串聯式質譜儀 ； 固相萃取法 ； 液相萃取法 ； 定量 ； QuEChERS

並列摘要

Heterocyclic aromatic amines (HCAs) comprise of a class of > 25 compounds, which primarily generated unintended hazardous substances by heating or processing of meats from cooking of proteinaceous foods at temperatures above 150℃. In addition, several of them have been found to be carcinogenic in animals, causing tumors in diverse organs across multiple species. The International Agency for Research on Cancer (IARC) has classified three HCAs (MeIQ, MeIQx, and PhIP) as being possible human carcinogens (Group 2B) and one HCA (IQ) to be a probable human carcinogen (Group 2A). In this study, two sample preparation strategies, liquid-liquid extraction (LLE) with solid-phase extraction (SPE) and quick, easy, cheap, effective, rugged, and safe extractions (QuEChERS) method, were investigated for the determination of 11 types of HCAs in meat products by LC-MS/MS. HCAs in sample were first extracted with acetonitrile by LLE, and followed by SPE using Oasis® MCX cartridges. In QuEChERS extraction, acetonitrile is used as LLME solvent, and PSA, C18EC and MgSO4 are served as dSPE sorbent. Both methods showed good performances on precision (RSD < 15.15%), accuracy (79.80-117.64%), recovery (52.39-116.88%), limit of quantitation for the spiked meat extract (0.01-10 ppb) and correlation coefficients (>0.993). QuEChERS extraction strategy gives better linear dynamic range and superior sensitivity in comparison with LLE-SPE approach. Eventually, HCAs were successfully quantified in real samples by two proposed approaches on LC-MS/MS system.

並列關鍵字

Heterocyclic aromatic amines ； HPLC-MS/MS ； QuEChERS ； SPE ； LLE ； quantitation

參考文獻

Skog, K. I., Johansson, M. A., and Jagerstad, M. I. (1998) Carcinogenic heterocyclic amines in model systems and cooked foods: a review on formation, occurrence and intake, Food Chem Toxicol 36, 879-896.

2. Doll, R., and Peto, R. (1981) The causes of cancer: quantitative estimates of avoidable risks of cancer in the United States today, Journal of the National Cancer Institute 66, 1191-1308.

3. Widmark, E. (1939) Presence of cancer-producing substances in roasted food, Nature 143, 984.

4. Nagao, M., Honda, M., Seino, Y., Yahagi, T., and Sugimura, T. (1977) Mutagenicities of smoke condensates and the charred surface of fish and meat, Cancer Letters 2, 221-226.

5. Sugimura, T., Nagao, M., Kawachi, T., Honda, M., Yahagi, T., Seino, Y., Sato, S., Matsukura, N., Matsushima, T., and Shirai, A. (1977) Mutagen-carcinogens in food, with special reference to highly mutagenic pyrolytic products in broiled foods, Origins of Human Cancer 4, 1561-1577.