檳榔花成分分析及其毒性評估

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Title:	檳榔花成分分析及其毒性評估 The compositions analysis and potential toxicities evaluation of the flower of Areca Catechu
Authors:	李昌澤
Contributors:	陳秋蘭嘉南藥理科技大學：藥物科技研究所
Keywords:	檳榔花急毒性基因毒性微核 micronuclei genotoxicity the flower of Areca catechu(FAC) acute toxicity
Date:	2010
Issue Date:	2010-12-30 15:33:56 (UTC+8)
Abstract:	前言：咀嚼檳榔嚼塊在台灣、印度、東南亞等許多國家都是很盛行的一種習慣。流行病學的研究已證實咀嚼檳榔嚼塊與口腔癌的發生有很高的相關性。而檳榔嚼塊中的檳榔鹼、檳榔多酚、銅離子、檳榔鹼相關的亞硝胺化合物及咀嚼時產生的活性氧化物等所造成的基因毒性、細胞毒性、及致突變性都被認為與口腔癌的發生有關。在台灣，檳榔的種植約佔所有農作物的7.25%，與檳榔有關的相關經濟產值更是佔所有農作物的第四名。檳榔花為檳榔樹的花穗，當檳榔園要廢園時，將其採下並作為食材入菜，但食用檳榔花後，是否有任何生理上的作用則不得而知。實驗目的：為了瞭解及預防檳榔花可能帶來的危害，因此本研究欲分析檳榔花內所含生物鹼及總多元酚的含量，並評估其在動物體內的急毒性、亞急毒性及基因毒性。實驗方法：本實驗分別以0.1%的醋酸或80%的丙酮萃取檳榔花內可能含有的生物鹼及總多元酚等物質後，以離子交換樹酯的層析管柱分離並用UV偵測其生物鹼含量。總多元酚則是以Folin-Ciocalteu的呈色反應來定量。急毒性試驗則是以胃管灌食大鼠最大劑量(2g/kg)的檳榔花萃取物後，連續觀察14天，並偵測其心跳、血壓及體重，同時收集血液分析餵食檳榔花萃取物後大鼠血中的生物鹼濃度，並比較餵食前後數值的變化。動物體內的基因毒性則是以微核試驗進行分析。亞急毒性試驗則是連續灌食28天SD大鼠低劑量的檳榔花醋酸或丙酮萃取物後，觀察其體重及微核數目。結果與討論：使用0.1%醋酸或80%丙酮能有效的萃取檳榔花裡的總多元酚及生物鹼。以0.1%醋酸萃取，每克新鮮檳榔花中含有0.23mg的檳榔鹼；而多酚類含量則為4.05mg。以80%丙酮萃取，每克新鮮檳榔花物中含有0.29g的檳榔鹼；而多酚類含量則為4.55mg。在急毒性方面，檳榔花萃取物並不影響大鼠的體重。用胃管餵食SD大鼠大劑量(2g/kg)的檳榔花醋酸萃取物後30分鐘，則會顯著增加雌鼠的心跳。而檳榔花的丙酮萃取物則稍微增加雌鼠的血壓；但反而降低雄鼠的血壓。在基因毒性方面，連續灌食ICR小鼠5天的檳榔花醋酸或丙酮萃取物，小鼠微核的數目都隨著劑量(1~4 g/kg) 的增加而顯著的增加，顯示大劑量的檳榔花萃取物連續食用可能具有基因毒性。在亞急毒性方面，連續28天灌食雌性SD大鼠低劑量(0.25與1g/kg) 的檳榔花醋酸或丙酮萃取物，並不影響大鼠的體重；而大鼠微核的數目則稍微增加，但並不具統計上的差異。這些結果顯示食用大量的檳榔花可能會影響心血管系統和造成基因的損傷。結論：因此建議食用前使用醋酸以去除出生物鹼及多酚類等可能的危害物質，進而減少食用檳榔花可能造成的風險。 Introduction: Betel quid (BQ) chewing is a popular oral habit in Taiwan, India, and many Southeast Asia countries. Epidemiological studies have correlated the BQ chewing with the development of oral cancer. The genotoxicity, cytotoxicity, and mutagenicity of BQ ingredients, including arecoline, areca polyphenol, copper, areca nut specific nitrosamines, and reactive oxygen species generated during chewing, are considered to be the major contributing factors to oral cancer. In Taiwan, the farmland for the betel palm (Areca catechu) represents 7.25% of total crop acreage and the betel nut industry there ranks fourth for production value among agricultural products. The flower of Areca catechu (FAC) is an inflorescence of the areca, locally consumed as vegetable when the betel palms fall into disuse. The physiological or toxic effects after consumption of this edible part of Areca catechu, however, have not been reported. Aim: The object of this study was to quantitate the contents of alkaloids and total phenolics in the FAC, then to estimate the acute toxicity, subacute toxicity, and genotoxicity in animals. Methods: The FAC was extracted by 0.1% acetic acid or 80% acetone, separately. Laboratory analysis of alkaloids was performed by HPLC with cation-exchange column and UV detector. The content of total phenolics in the extracts of FAC was determined spectrometrically according to the Folin-Ciocalteu procedure. To evaluation the acute toxicity, the SD rats were oral gavage with a single large dose (2 g extracts/kg body weight) of FAC, and then the body weights and the cardiovascular parameters were estimated at the 7th and 14th day. The genotoxicity of the FAC extracts was estimated by micronuclei test in ICR mice. In the subacute toxicity, the body weights and the micronuclei were estimated when oral gavage the FAC extracts with a repeated low doses for 28 days in SD rats. Results and discussions: These results suggest that the alkaloids and total phenolics were efficiency extracted by 0.1% acetic acid or 80% acetone. The arecoline contents were 0.23 mg/g FAC extracted by 0.1% acetic acid and 0.29 mg/g FAC extracted by 80% acetone, separately. The total phenolics were 4.05mg/g FAC extracted by 0.1% acetic acid and 4.55 mg/g FAC extracted by 80% acetone, separately. In a single high dose (2g/kg) oral gavage, 0.1% acetic acid FAC extracts increased the heart rate in female SD rats, but 80% acetone FAC extracts decreased the blood pressure in male SD rats. In the genotoxicity, 0.1% acetate FAC extracts induced micronuclei in a dose-dependant manner. In the subacute toxicity, low dose (0.25~1g/kg) of FAC extracts increases the micronuclei numbers in female SD rats, but has not significant difference. These results suggest that ingestion of large amount of FAC may affect the cardiovascular system and cause genetic damage. Conclusions: The study indicated that pretreatment of acetic acid might extract the arecoline and polyphenolics, then consequently reduce the risk of ingestion of FAC.
Relation:	校內馬上公開，校外一年後公開，學年度：98，53頁
Appears in Collections:	[Dept. of Pharmacy] Dissertations and Theses

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