蘇丹紅 III 檢驗方法開發暨秀麗隱桿線蟲試驗

參考 TFDA公告方法，開發 HPLC 及拉曼光譜蘇丹紅 III 分析方法，並檢測市售辣椒油、辣椒粉，藉由蘇丹紅染料為脂溶性之特性，溶於油中，經正己烷震盪萃取 20 分鐘後，靜置 30 分鐘，取上清液於 50℃濃縮機濃縮，經過 0.45 μm PVDF 過濾膜過濾後，檢液待上機分析；分析條件移動相 A 1%甲酸水溶液，移動相 B 乙腈，於 500 nm 下檢測，進行 37 分鐘梯度程式沖提，圖譜出峰時間與標準品相比較，發現市售辣椒油、辣椒粉成分天然，皆無被摻假之疑慮，且以拉曼光譜佐證；蘇丹紅 III 分析方法之確效試驗，配製序列濃度製作檢量線，線性範圍為0.8 μg/mL ~100 μg/mL之間，檢量線之相關係數(r)均大於 0.99，並進行同日內 (Intra-day)、異日間(Inter-day)試驗，同日內(Intra-day)之標準偏差 R.S.D. (Related Standard Deviation 為 0.004%〜0.028%；在異日間(Inter-day)試驗為 0.005%〜0.021%，均小於 2%，取檢品溶液添加低、中、高濃度之標準品，進行回收率(Recovery)試驗，結果顯示 Sudan III 回收率為 98%〜 112%之間，顯示此 HPLC 分析方法，具有良好的適應性(System Suitability)、精密度(Precision)、準確度(Accuracy)。拉曼光譜分析測試，藉由樣品中特殊官能基訊號，作為進一步快篩鑑別；秀麗隱桿線蟲毒性試驗顯示，10 ppm Sudan III 樣品，測試 1 天，足以造成秀麗隱桿線蟲體內因氧化壓力產生綠色螢光，且隨著時間變化 0 分鐘至 120 分鐘，綠色螢光逐漸明顯，直到秀麗隱桿線蟲死亡而產生藍色螢光。

關鍵字

蘇丹紅染料；脂溶性偶氮染料；毒化物；秀麗隱桿線蟲

並列摘要

Taking into account the TFDA public experimental method, an HPLC method was developed and the Raman spectroscopy was also used for Sudan III analysis to check whether the chili oil and chili powder have contents of Sudan III. Sudan dyes are fat soluble and therefore was dissolved in oil. After 20 minutes of shaking extraction with n-hexane, the mixture was allowed to stand for 30 minutes, and the supernatant was concentrated at 50 ° C. Then filtered through a 0.45μm PVDF filter membrane and the solution was analyzed in HPLC. The HPLC analytical conditions include mobile phase A which was treated with 1% formic acid aqueous solution, and mobile phase B treated with acetonitrile. Detection was at 500 nm wavelength, and the gradient was programmed for 37 minutes. The peak time of the sample was compared with the standard product, considering that purchased chili oil and chili powder ingredients are naturally. About the determination test of Sudan III analysis method, preparation of sequence concentration was performed to make a calibration curve with a linear range of 0.8 μg/mL ~ 100 μg/mL. The correlation coefficient (r) of the calibration curve is greater than 0.99, and the intra-day and inter-day tests are performed. The Related Standard Deviation standard of deviation the intra-day is 0.004 %~0.028%. In the inter-day test, 0.005%~0.021% was obtained. Both of these results are less than 2%. The test solution was then added with low, medium and high concentration standards and the recovery test was carried out. The recovery rate of Sudan III is between 98% and 112%, indicating that this HPLC analysis method has good system suitability, precision and accuracy. Raman spectroscopy test, with a special functional signal in the sample was used as a further fast screening identification. C. elegans toxicity test showed that a low concentration of Sudan III sample was 10 ppm enough to cause green fluorescence in the C. elegans due to oxidative stress. Within a period from 0 minutes to 120 minutes with time green fluorescence gradually became apparent until the C. elegans died and blue fluorescence was produced.