臺灣博碩士論文加值系統

蘆薈被廣泛使用在健康飲品、外用藥膏、洗護用品和化妝品上，其生理活性物質包含蘆薈大黃素、皂素以及蘆薈素等，而其中的蘆薈素具有抗氧化能力，且有致瀉作用。國內飲料店與超市提供各式各樣的蘆薈飲料，其中尤以蜂蜜蘆薈汁最受消費者喜愛，因蘆薈飲料之研究闕如，故擬針對市面上手搖飲料店、超市或賣場含蘆薈之飲品，進行蘆薈素含量之調查。首先探討蘆薈素的萃取回收率，結果顯示蘆薈素萃取以甲醇做為溶劑、pH 值4為較佳條件，萃取液經離心過濾後以液相層析串聯質譜儀 (Liquid chromatography tandem mass, LC-MS/MS) 進行蘆薈素檢測。質譜儀設定程式以417 m/z為母離子， 417 > 297 m/z 為定量離子，417 > 268 m/z、 417 > 251 m/z為定性離子，蘆薈素aloin A和 aloin B 於 0.364 ppb ~ 0.675 ppm 範圍內之線性迴歸係數分別為 0.9996 和 0.9996。添加平均回收率介於 71.4 ~ 103.5% 之間，經內部標準品修正之回收率介於 81.8 ~ 116.7%，蘆薈素之同日間及異日間分析之變異係數 (CV%) 小於 2.48%，aloin A定量極限為 0.33 ppb，而aloin B 定量極限為 0.18 ppb。LC-MS/MS以分子量做為定性與定量分析具專一性，分析一件蘆薈素樣品所需時間為12分鐘，優於以HPLC-DAD分析之80分鐘，且可得更低的偵測極限與定量極限。以本研究建立之LC-MS/MS方法檢驗飲料蘆薈素之含量，測試41件蘆薈飲料樣品，pH值為3.02 ~ 4.60，其中6件未檢出蘆薈素，而其他樣品檢出濃度範圍在0.005 ~ 2.921 mg/kg之間，依衛生福利部規定飲料中蘆薈素的限量標準為 0.1 mg/kg，則超標樣品數量為17件

Aloe has been extensively used for health drinks, external ointments, toiletries and cosmetics. Its physiologically active substances included aloe emodin, saponin and aloin, of which aloin possessed antioxidant capacity and diarrhea effect. The domestic beverage shops and supermarkets offer a variety of aloe drinks and related products, of which the product aloe juice with honey is the most popular. Little information of aloe drinks has been available. The purpose of this study was to investigate the contents of aloin in aloe drinks and related products purchased from beverage stores and supermarkets. First, the extraction recovery of aloin was studied. Results showed that methanol as solvent, pH value at 4 were better conditions for aloin extraction. After centrifugation and filtration, the extractive solution was directly subjected to LC-MS/MS (Liquid chromatography tandem mass) to determine the amount of aloin. The mass spectrometer was programmed to monitor the precursor ion at m/z 417 and the product ion at m/z 297, m/z 268, m/z 251. The results showed the liner range of 0.364 ppb ~ 0.675 ppm could be achieved, and correlation coefficient (R2) of aloin A was 0.9996 and aloin B 0.9996. The spike recoveries were 71.4 ~ 103.5%, and the spike recoveries with internal standard were 81.8 ~ 116.7%. The coefficient of variation (CV%) of aloin was less than 2.48% for intra-day and inter-day detection. The limit of quantification (LOQ) with aloin A and aloin B were 0.33 ppb and 0.18 ppb, respectively. By monitoring molecular weight, LC-MS/MS has specificity for qualitative and quantitative analysis. It took 12 min to analyze a sample, more time-saving than 80 min by HPLC - DAD. Besides, lower LOQ and limit of detection (LOD) of aloin were found by LC-MS/MS method. The present method has been successfully applied to the analysis of aloin in aloe drinks and related products. The 41 aloe drink samples with pH value of 3.02 ~ 4.60 were investigated. Six samples were found no aloin, while others with the concentration ranged from 0.005 ~ 2.921 mg/kg. According to the Ministry of Health and Welfare, the sanitation limit of aloin content in beverage is 0.1 mg/kg. Results showed that 17 samples investigated do not meet the requirement of this limit.

目錄
中文摘要 …..Ⅰ
英文摘要 …..Ⅱ
目錄 …..Ⅲ
表目錄 …..Ⅴ
圖目錄 …..Ⅵ
附表目錄 …..Ⅶ
附圖目錄 …..Ⅷ
壹、前言 1
貳、文獻整理 2
一、 蘆薈介紹 2
(一) 蘆薈的構造 2
(二) 蘆薈品種介紹 2
二、 蘆薈之化學成分與營養物質 3
(一) 蘆薈果肉與分泌液之化學成分 3
(二) 蘆薈凝膠之化學成分 3
(三) 蘆薈素 3
三、 蘆薈生理活性物質 4
四、 蘆薈之抗菌活性 4
五、蘆薈生理活性物質萃取與分析 5
六、蘆薈萃取液對糖尿病大鼠的抗氧化活性 6
七、蘆薈萃取物的急性毒性與致瀉作用 6
八、液相層析質譜儀介紹 7
参、材料與方法 8
一、 實驗材料 8
(一) 蘆薈飲品 8
(二) 標準品 8
(三) 藥品材料 8
(四) 層析管柱 8
(五) 儀器與裝置設備 8
二、 實驗流程 10
三、 實驗方法 11
(一) 蘆薈樣品之製備 11
(二) 蘆薈素萃取溶劑、p H 值與溫度之測試 11
(三) LC-MS/MS分析方法之建立 11
1. 儀器參數設定 11
2. 檢量線之製作 12
3. 管柱之選擇 13
4. 移動相之條件 13
5. 再現性分析 14
(四) CNS方法15502之檢測 14
1. 前處理方法 (經濟部標準檢驗局，2011) 14
2. 儀器分析條件 14
3. 鑑別試驗及含量測定 15
肆、結果與討論 16
一、蘆薈素萃取最適溶劑、pH值與溫度之探討 16
二、LC-MS/MS檢測蘆薈素方法之建立 16
(一) 儀器參數設定 16
(二) 檢量線之製作 17
(三) 再現性分析 17
三、CNS方法與LC-MS/MS檢測蘆薈素之比較 17
四、市售蘆薈飲品之蘆薈素含量調查 18
伍、結論 20
陸、參考文獻 21

表目錄
表 一、萃取溶劑與pH 值對蘆薈素萃取回收率之影響 24
表 二、 蘆薈素以 LC-MS/MS 檢測之濃度範圍、相關係數與線性迴歸 25
表 三、以 LC-MS/MS檢測同日間 (Intra- Day) 及異日間 (Inter- Day) 再現性分析 26
表 四、 蘆薈素以 HPLC-DAD 檢測之濃度範圍、相關係數與線性迴歸 27
表 五、 HPLC-DAD 與 LC-MS/MS 檢測蘆薈素之偵測極限及定量極限 28
表 六、 HPLC-DAD 與 LC-MS/MS 檢測蘆薈素及內部標準品之滯留時間 29
表 七、市售蘆薈飲品之樣品資訊及 pH 值 30
表 八、包裝蘆薈飲品與蘆薈健康飲品之樣品資訊及 pH 值 31
表 九、飲料店蘆薈飲料之蘆薈素含量調查 32
表 十、市售包裝蘆薈飲料之蘆薈素含量調查 33
表 十一、市售包裝蘆薈保健飲料之蘆薈素含量調查 34

圖目錄
圖 一、加熱溫度與 pH 值對蘆薈素萃取回收率之影響 35
圖 二、加熱溫度與 pH 值對蘆薈素萃取回收率之影響 36
圖 三、蘆薈素 ESI (負電荷) 全掃描之離子圖 37
圖 四、蘆薈素標準品離子光譜圖(負電荷模式) 38
圖 五、內部標準品離子光譜圖(負電荷模式) 39
圖 六、蘆薈素 ESI (正電荷) 全掃描之離子圖 40
圖 七、樣品 LC-MS/MS 層析圖與離子光譜圖 41
圖 八、蘆薈素 aloin A 以 LC-MS/MS 檢測之線性迴歸 42
圖 九、蘆薈素 aloin B 以 LC-MS/MS 檢測之線性迴歸 43
圖 十、蘆薈素內部標準品以 LC-MS/MS 檢測之線性迴歸 44
圖 十一、蘆薈素 aloin A (a) 與 aloin B (b) 以 HPLC 檢測之線性迴歸 45
圖 十二、Aloin A 於 LC-MS/MS 的偵測極限 46
圖 十三、Aloin B 於 LC-MS/MS 的偵測極限 47
圖 十四、Aloin A 於 LC-MS/MS 的定量極限 48
圖 十五、Aloin B 於 LC-MS/MS 的定量極限 49
圖 十六、蘆薈素 1 ppm、3 ppm 及 10 ppm 之 HPLC-DAD 層析圖 50
圖 十七、蘆薈素 (10 ppm) 之光譜圖 51
圖 十八、蘆薈素及其內部標準品以 LC-MS/MS 分析之層析圖 52

附表目錄
附表 一、南非蘆薈品種保存狀態與用途 53
附表 二、蘆薈果肉與分泌液植物化學成分 61
 
附圖目錄
附圖 一、蘆薈葉剖面結構圖 62
附圖 二、蘆薈凝膠的化學成分 63
附圖 三、蘆薈生理活性物質結構圖 64
附圖 四、皂素結構圖 65

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