利用高效液相層析法檢測膠囊錠狀食品中葉黃素及玉米黃素之含量

Determination of Lutein and Zeaxanthin Contents in Capsules or Tablets by a HPLC Method

陳瑋芸(Wei-Yun Chen) ； 蔡沁玹(Ching-Hsuan Tsai) ； 江峻蔚(Chun-Wei Chiang) ； 吳白玟(Pai-Wen Wu) ； 曾素香(Su-Hsiang Tseng) ； 高雅敏(Ya-Min Kao) ； 闕麗卿(Li-Ching Chiue) ； 施養志(Yang-Chih Shih)

《食品藥物研究年報》 5期 (2014/12) Pp. 17-25

摘要

葉黃素及玉米黃素可避免老年性黃斑病變(age-related macular degeneration,AMD)。在調查市售膠囊錠狀食品中葉黃素及玉米黃素之含量時，為能提高檢驗效率並同時分析二種待測物，本研究擬建立以高效液相層析儀(high performance liquidchromatograph, HPLC)搭配光二極體陣列檢出器(photodiode array detector)同步分析膠囊錠狀食品中葉黃素及玉米黃素含量之檢驗方法。檢體以丙酮：去離子水溶液超音波振盪萃取，加入20%氫氧化鉀酒精溶液進行皂化，經0.2% BHT之乙醚-乙酸乙酯溶液進行液/液萃取，合併上層液經減壓濃縮後，殘留物以乙醇-乙酸乙酯溶液溶解並定容，再以HPLC進行分析。使用之層析管柱為Ascentis® RP-Amide，移動相溶液為乙腈與去離子水以95:5 (v/v)之比例混合，流速為1 mL/min，檢測波長為450 nm。經實驗分析後，葉黃素及玉米黃素標準曲線之線性範圍均為0.1-100 μg/mL，線性回歸方程式之判定係數(R^2)分別為0.9993及0.9959。接著進行添加回收試驗，將已知濃度的標準品添加至粉狀基質及沙拉油基質後，求得葉黃素及玉米黃素於粉狀基質之回收率介於77.4-95.4%之間，添加回收率之相對標準偏差則介於3.2-6.0%之間；葉黃素及玉米黃素於沙拉油基質之回收率介於86.1-124.6%之間，添加回收率之相對標準偏差則介於8.5-11.1%之間。進一步分析葉黃素及玉米黃素之定量極限濃度，由實驗結果得知葉黃素及玉米黃素於粉狀基質之定量極限為125 mg/kg，於沙拉油基質之定量極限為625 mg/kg。

關鍵字

膠囊錠狀食品 ； 酯化型葉黃素 ； 酯化型玉米黃素 ； 皂化 ； 高效液相層析法

並列摘要

Lutein and zeaxanthin can reduce the risk of Age-related Macular Degeneration (AMD). This study was aimed to develop a analytical method for lutein and zeaxanthin contents in capsules or tablets by HPLC-DAD at the same time. The homogenized samples were sonicated in acetone solution, and then 20% ethanolic KOH saponification agent was added. After saponification, samples were extracted three times using ether/ethyl acetate (1:1, v/v) solution containing 0.2% BHT. The supernatants were collected and combined, then evaporated to dryness. After evaporation, the oily residue was completely dissolved with ethanol/ethyl acetate (1:1, v/v) solution contain 2% BHT then transferred to a 25 mL volumetric flask and made up to the volume. The aliquot was filtered and injected to a HPLC. The HPLC was equipped with a Ascentis® RP-Amide column using ACN and water (95:5, v/v) as the mobile phase at a flow rate of 1 mL/min. The chromatography was monitored by absorbance at 450 nm. The linear coefficients of regression equation of lutein and zeaxanthin were 0.9993 and 0.9995, respectively. Recovery analysis was performed by spiking standard compounds into blank powder and oil materials. The recoveries of lutein and zeaxanthin were from 77.4 to 95.4% and the coefficients of variation were from 3.2 to 6.0% for powder material. The recoveries of lutein and zeaxanthin were from 86.1 to 124.6% and the coefficients of variation were from 8.5 to 11.1% for oil material. The quantitation limits of lutein and zeaxanthin were 125 mg/kg for powder material and 625 mg/kg for oil material.

並列關鍵字

capsule ； tablets ； lutein ester ； zeaxanthin easter ； saponification ； HPLC