利用奈微米化技術開發多功能組合性保健食品之研究－基因毒理及食用安全性評估

Abstract

近年來，在發展保健食品過程，相當注重協力作用(synergy)之觀念，嘗試結合多種食材開發「組合性」的配方產品(complex formulas)，即將多種明確已知活性成分資料的食材相互配合使用，結合多種保健食材和成份的生理活性和優點，達到「多功能」的目的。本群體計畫乃依此概念分別組合具預防代謝症候群相關疾病、調節免疫功能、抗氧化等多種生理功能保健食品配方，第一階段為A, B, C配方，第二階段為I及K配方，第三階段為New J 配方。唯當一個新興食品上市前，評估潛在毒性之風險與食用安全性不容忽視。依照我國衛生署訂出的健康食品安全性評估方法，分別評估基因毒理試驗與動物毒性試驗。基因毒理包括：沙門氏菌致突變性(安姆氏試驗)、中國倉鼠卵巢細胞染色體變異及小鼠紅血球微核等測試。結果顯示第一階段三種配方(A, B, C)並無基因毒性作用，同時對小鼠之口服LD50值大於5 g/kg body weight。唯配方C小鼠血清BUN及creatinine值均較對照組偏高，檢視配方C組成中含有冬青葉，可能具腎毒性，建議抽換冬青葉，再進行新組合配方之安全性評估。第二階段保健食品(I, K)進行相同基因毒理試驗，結果顯示並無基因毒性作用。第三階段最後配方(New J)進行安姆氏試驗確定並無基因毒性。在食用安全性評估之急性毒性試驗結果顯示，New J 對大鼠口服之LD50值為大於5 g/kg body weight；28天亞急毒性試驗，則分別給予低(1 g/kg)、中(3 g/kg)及高(5 g/kg)三種劑量連續餵食大鼠28天，分析體重、飼料消耗量、血液學、血液生化學、尿液學及病理學等各項檢查，結果顯示，在中、高劑量組之雄鼠與雌鼠之體重與飼料消耗量均有下降趨勢，中及高劑量組之雌鼠血清白蛋白、白蛋白/球蛋白比率、鹼性磷酸酶、澱粉酶、天門冬酸轉氨酶、氨基丙酸轉氨酶、肌酸激酶、葡萄糖、總膽紅素、總膽固醇、三酸甘油酯、鈣、磷、氯、鉀、鎂和鈉等數值與對照組雖有顯著差異(p<0.05)，但仍在正常生理值範圍內，體內重要器官均無明顯肉眼及組織病理變化。綜合以上結果，配方New J 對大鼠之28天餵食無毒害作用劑量 (no-observed-adverse-effect-level, NOAEL) 為1 g/kg bw/day。

In the recent global market of functional foods, the concepts of “synergy” and “multifunctional” were popular and have driven the development of complex formulas using multi-ingredients. Several complex formulas with or without micronization treatments will be developed using different natural plants and herbal materials with the goal to prevent diseases relating to the aging, metabolism, various degenerative disorders and immunomodulatory problems. As new material come on the market, their genotoxicity and safety should be evaluated first. The aim of this study was to evaluate the possible genotoxicity of three different stages health food formulas following Ames test, chromosome aberration, and micronucleus tests and animal feeding toicity. Results revealed that all formulas (A, B, C, I, K) had no mutagenicity. Additionally, the acute oral LD50 of formula A, B and C in mice was greater than 5 g/kg body weight. However, significant increase of BUN level was noted in the formula C treated group. The of formula C contained green mate leaf (Ilex paraguariensis) that had genotoxicity and associated with the risk of renal cell carcinoma in human. For the safety, it was suggested that the green mate leaf need to exclude in the next new formulation. On second stage, no mutagenicity of I and K formulas was found. Furthermore, the final product of New J formula was evaluated by Ames and animal toxicity tests. Rat were daily administered (by gavage) at doses of 1, 3 and 5 g/kg body weight for 28 days. Results revealed that no mutagenicity of New J and the acute oral toxicity was greater than 5 g/kg bw. In the 28-day feeding toxicity, all animals survived and no clinical signs were observed. However, significant different parameters in the middle and high dose groups of male rats were noted, induding body weight, body weight gain and feed consumption. It was alo noted significant difference in the middle and high-dose of female rats in blood biochemistry values when compared with control rats. Based on the result, the no observed-adverse-effect level (NOAEL) of New J formula is 1 g/kg bw/day in the 28-day feeding toxicity in rats.