分析技術開發及應用研究: Part I.自動瞬時量測空氣與水中微量揮發性有機物之分析研究; Part II.以高壓萃取法萃取斑蟊與芭樂葉中有效成分及生物活性之研究

Development and Application of Analytical Technique: Part I. The Studies of Automatic In-situ Monitoring of Volatile Organic Compounds in the Ambient and Wastewater; Part II. The Studies of Bioactive Compounds of Mylabris and Guava Leaves Extracted by Pressurized Extraction

粘弘吉(Nian, Hungchi)

指導教授 ： 羅俊光 ； 瞿港華

國立清華大學/原子科學院/生醫工程與環境科學系/博士(2009年)

摘要

本研究論文為分析技術開發及應用之研究，共分為兩部份：第一部份--自動瞬時量測空氣與水中微量揮發性有機物之分析研究。本研究空氣監測部分利用實驗室開發完成的自動化氣相層析質譜儀，建立自動採樣分析之氣相層析質譜系統，進行連續十一天之環境監測，期間納莉颱風侵襲本島，儀器仍持續正常運作。颱風滯留三天裡，量測丙酮濃度低至偵測極限以下；相對極性較低之物種而言，甲苯平均濃度值為3.0 ppbv。得知極性高之物質容易隨著雨水沖刷而流至地面，雨量之多寡將對極性物種與非極性物種之量測有著不同程度影響。
另外，本研究水體監測部份針對經污水處理廠處理後廢水中揮發性有機化合物進行監測系統建立及分析，研究中監測系統發展為完全自動進樣、自動化的分析系統，有效縮短分析時間且運用在現場即時監測，有效取得24小時連續即時的分析結果並節省現場採樣、人力分析與資源的運用。
第二部份--以高壓萃取法萃取斑蟊與芭樂葉中有效成分及生物活性之研究。在斑蝥素的萃取上， CO2搭配乙腈約萃取出9.18 mg/g斑蝥素。液態二氧化碳萃取斑蝥素的與傳統的萃取方法有較明顯的差別，雖說效果並非最佳的萃取方法，但其主要優點是可以減少有機溶劑的使用，縮短萃取時間(液態二氧化碳30min;傳統萃取需65小時)並且幾乎不需要再經由其他純化的步驟就可以得到斑蝥素。
在芭樂葉萃取方法中，以高壓甲醇萃取法有較好的總類黃酮萃取效率，而其中利用加壓萃取不但省時而且省溶劑，所以實驗的萃取方式採用高壓甲醇萃取法。在動物餵食測試推論出quercetin對於芭樂葉降血糖的能力有一定的幫助，但是並非芭樂葉降血糖主要的有效成分，應該還有其他更重要的活性成分導致在動物實驗中明顯的降血糖效果。

關鍵字

揮發性有機物 ； 瞬時量測 ； 高壓萃取法 ； 斑蟊 ； 芭樂

並列摘要

The thesis contained two parts. In the first part, automatic in-situ monitoring systems of volatile organic compounds (VOCs) in the ambient and wastewater were developed.The study describes continual monitoring of the volatile organic air pollutants, acetone and toluene, in HsinChu Science Park (HSP) during an inclement weather occurrence, i.e., typhoon. A total of 53 polar and nonpolar compounds were identified and analyzed. The method detection limit (MDL) was 0.4 ppbv on average. The study presents a novel system, which continuously monitors polar VOCs in industrial wastewater. The system utilizes an ultrasound coupled autosampler, headspace solid-phase microextraction (HS-SPME), and a gas chromatography/flame ionization detection GC/FID for onsite, real-time detection and quantification of VOCs. The MDL was between 3.19ppb and 7.57ppb.

In another part, the study using pressurized liquid carbon dioxide to extract cantharidin from Mylabris was investigated. The result achieves green extraction of bioactive ingredients in the insect, and the extraction efficiency of 95% (three times) was obtained at pressure of 1 MPa, temperature of 60°C, static extraction time of 30 minutes and with 4 ml of acetonitrile as modifier. In addition to the identification of the extract, i.e., cantharidin, by chromatographic approach, an in vitro assay for the effect of the extract on the cell was performed, the result reveals that the cantharidin-extracted is efficacious for primary medicinal available.
The study proposes to verify the hypoglycemic effect of the flavonoids extracted from guava leaves on type 2 diabetic mice. When comparing the total flavonoid contents from various extraction methods, pressurized-methanol extraction had the highest levels (38.4±1.5 mg g-1). The major flavonoids of guava leaves were analyzed with mass spectrometry (MS) coupled with a high-performance liquid chromatograph, diode-array detector (HPLC-DAD). To confirm the hypoglycemic effect, an animal fasting test was conducted. In summary, both primary extracts (dose of 0.5 g Kg-1) and isolated flavonoids (dose of 40 mg Kg-1) from guava leaves had hypoglycemic effects (P<0.001). However, the quercetin flavonoid in guava leaves was only one of the active compounds with hypoglycemic activity, not the most major active compound.

參考文獻

27. 黃敬德、謝有容，”固相微萃取技術之原理及其應用”，1998，Chemistry, Dec.

4. U.S. EPA. Compendium method TO-16, Long-path open-path Fourier transform infrared monitoring of atmospheric gases; 1996.

5. U.S. EPA. Compendium method TO-17, The determination of volatile organic compounds (VOCs) in air using active sampling onto sorbent tubes; 1997.

13. Magdalena Ligor, Boguslaw Buszewski,“Determination of menthol and menthone in food and pharmaceutical products by solid-phase microextraction–gas chromatography”Journal of Chromatography A 1999, 847,161-169

14. Beatriz Cancho, Francesc Ventura, M Teresa Galceran,“Determination of aldehydes in drinking water using pentafluorobenzylhydroxylamine derivatization and solid-phase microextraction”Journal of Chromatography A 2001, 43,1-13