利用新穎生物快速篩檢法檢測台灣土壤與底泥樣本中戴奧辛濃度

The study of a new fast-screen dioxins bioassay applied in Taiwanese contaminated samples in soil and sediment

郭彥祐(Yan-You Gou)

指導教授 ： 趙浩然

國立屏東科技大學/工學院/環境工程與科學系所/碩士(2011年)

https://doi.org/10.6346/NPUST.2011.00154

摘要

近年來戴奧辛污染事件層出不窮，如果僅僅依靠傳統分析戴奧辛方法(GC-HRMS)以時效性層面上看來是不足的，因此當污染事件發生時如能先進行初步篩測，而後進一步利用化學分析檢測，如此即能解決時效性問題，本研究利用腺病毒載體(Adenovirus)進行CALUX方法改良，將此方法定義為DRE-driven luciferase assay。利用此方法進行2,3,7,8-TCDD標準品、液態標準品、環境基質(CRM)、土壤、底泥樣本測試。本次研究方法之偵測極限值為0.209 pM，土壤樣本預估值與GC-HRMS兩者數據分別取對數轉換後進行一次線性迴歸，兩者具有良好的關聯性(R2=0.725)。底泥樣本預估值與GC-HRMS同樣也具有良好的關聯性(R2=0.787)。將土壤與底泥樣本進行單一樣本T檢定後發現呈現偽陽性與偽陰性數據有達到顯著差異(p<0.001)，同時將檢測土壤數據進行分組，利用Mann-Whitney U tests檢定發現，當樣本冷光讀值落在EC50以下區段時與GC-HRMS測值具有良好的關聯性(平均比值為2.07倍)，有達到顯著差異(p=0.006)，同時本研究也發現當標準品EC50冷光讀值為10000 RLU以下時底泥樣本預估值與GC-HRMS測值平均比值為3.26倍，與EC50冷光讀值高於10001 RLU兩者數據達到顯著的差異(p<0.001)。因此由上述數據顯示本實驗方法應可適用於真實土壤及底泥樣本快速篩測工具，數據均呈偽陽性，且如果能夠使樣本冷光讀值落在EC50以下區段與標準品EC50數據低於10000 RLU時，本實驗方法預測值能夠更加準確。

關鍵字

生物快速篩檢戴奧辛方法 ； 土壤 ； 底泥 ； DRE-driven luciferase assay

並列摘要

While dioxins pollution events have been frequently occurred, a rapid and cost-effective screening method of dioxin in Taiwan is relatively urgent. This study investigated dioxins contamination in Taiwanese environmental samples from soils and sediments by analyzing DRE-driven luciferase assay. DRE-driven luciferase assay is a bioassay with H4IIE cell lines transfected by DRE-driven adenovirus. Associations of analytical outcomes between high-resolution gas chromatography/high-resolution mass spectrometry (GC-HRMS) and DRE-driven luciferase assay were examine by chemical standards and environmental samples including 2, 3, 7, 8-TCDD, liquid standards (EDF-5416, 5417), certified reference material (CRM), soil, and sediment.. Owing to determination of working criteria in a DRE-driven luciferase assay, the results of this bioassay are categorized and quantified to find their optimum by several statistical methods Limit of detection in this DRE-driven luciferase assay was 0.209 pg-ADL-TEQ/mL. After fitting into the Hill equation, the regression curve of the soil and sediment was shown with R2 = 0.725, and 0.787, respectively. This study found a significant difference between false negative and false positive in soil and sediment (p<0.006) by one-sample T-test. A Mann-Whitney U test was conducted to evaluate the hypothesis that ratio (GC-HRMS assay/DRE-driven luciferase assay) was more nearly 1 when luciferase value was under EC50 as opposed to above EC50 (p=0.006). While EC50 was lower than 10000 RLU, the ratio was also more nearly 1. According to the results, the DRE-driven luciferase assay probably has the potential to be a powerful technique for pre-screening dioxin samples from different sources, including soils and sediments.

並列關鍵字

bioassay ； soil ； sediment ； DRE-driven luciferase assay

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