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Title: 鯖魚油的萃取及利用基質輔助雷射脫附游離飛行時間質譜法對其三酸甘油酯的分析
Extraction of Mackerel Oil and Analysis of its Triacylglycerol by Matrix-assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry
Authors: Wang, Yu-Fen
Contributors: NTOU:Department of Food Science
Keywords: 鯖魚油;溶劑萃取法;索式萃取法;微波輔助萃取;MALDI-TOF MS;分析條件;三酸甘油酯;磷脂醯膽鹼
mackerel oil;Bligh and Dyer;soxhlet extraction;microwave assisted extraction;MALDI-TOF MS;analysis condition;triacylglycerol;phosphatidylcholine
Date: 2019
Issue Date: 2020-07-03T08:43:50Z
Abstract: 魚油對人類健康具重要性,常製成膳食補充劑,但市場上仍有摻偽現象。魚油容易因外在環境氧化裂解而導致標示不符。鯖魚分布廣、廣受人類喜愛,可製作為膳食補充劑。基質輔助雷射脫附游離-飛行時間質譜法具有樣品製備簡單、質譜分析過程快速的優點。本篇研究探討鯖魚油的不同萃取條件,使用溶劑萃取法、索式萃取法和微波輔助萃取法。微波輔助萃取鯖魚用乙醇做溶劑在微波功率時間 300 W/30 sec (14.3%) 時相對於傳統溶劑萃取 (10.3%) 和索式萃取 (3%) 有更高萃取率,取得油與極性萃取物。探討 MALDI-TOF MS 分析鯖魚油三酸甘油酯 (TAG) 時之最佳條件,使用不同基質、雷射強度、樣品基質比例和點樣次數,發現使用 2,5-dihydroxybenzoic acid 做基質、雷射強度比例 90%、樣品基質比例 (1/1, v/v) 和點樣三次時,可達到最佳的分析效果。使用 MALDI-TOF MS 分析鯖魚油之 TAG 將其質譜數據與線上資料庫 LIPID MAPS Online Tools 比對後,鑑別出鯖魚油中的 25 種 TAG 分子和推測出其中的脂肪酸組成,有 m/z 799.7 (C46:1, MPP1)、801.7 (C46:0, MPP)、813.6 (C48:8)、815.6 (C48:7, LM1D6/M1M1E5)、817.6 (C48:6, LP1E5/LLn3Ln3)、819.6 (C48:5, MME5)、825.7 (C48:2, MPL2/PP1P1)、827.7 (C48:1, PPP1/MPO1)、829.7 (C48:0, PPP)、839.6 (C50:9)、841.6 (C50:8, M1M1D6/LLn3E5)、843.6 (C50:7, LP1D6/M1P1E5)、845.7 (C50:6, MMD6/MP1E5)、849.7 (C50:4, ML2L2)、851.7 (C50:3, PPLn3/PP1L2)、853.7 (C50:2, PPL2/PP1O1)、867.6 (C52:9, LLn3D6/M1Ln3E5)、869.7 (C52:8, M1P1D6/MLn3E5)、875.7 (C52:5, PPE5/PL2Ln3)、877.7 (C52:4, PL2L2)、879.7 (C52:3, PO1L2)、889.6 (C54:12)、891.6 (C54:11, LE5D6/M1E5E5)、913.6 (C54:0)、915.6 (C56:13)。將其與鯡魚油和鱈魚肝油 之 TAG 對比,m/z 813.6、815.6、839.6、843.6、889.6、891.6 未存在於其中,m/z 799.7、825.7、827.7、829.7、845.7、849.7、853.7、869.7、875.7、877.7、879.7 為共同存在於鯡魚油和鱈魚肝油中。將其與大豆油、葵花油等植物油中共有的 TAG 對比具有顯著差異性。顯示測得之鯖魚油 TAG 分子對於鯖魚相關產品或研究做物種鑑定和鑑別在魚油或魚油飼料中加入較低價植物油的摻偽產品之潛力。同時,鯖魚油經 MALDI-TOF MS 分析後可測得其中的磷脂醯膽鹼 (PC),分別是 m/z 803.6、805.6、831.7、833.7,分子組成依序為 PC(18:2/20:5)、PC(16:0/22:6)、PC(18:1/22:6)、PC(20:1/22:6)。
Fish oil is used as a dietary supplement for human health, but it has often been adulterated on the market for years. Fish oil is easily degraded by the environmental factors, leading to mislabel or insecurity. Mackerel is widely distributed and its oil has been made into dietary supplements. Matrix-assisted laser desorption-time-of-flight mass spectrometry (MALDI-TOF MS) has the advantage of easy sample preparation and rapid mass spectrometry analysis. This study explores the optimum extraction conditions for mackerel oil using Bligh and Dyer, soxhlet extraction and microwave-assisted extraction. It was found that the use of microwave-assisted extraction of mackerel oil at a microwave power of 300 W for 30 sec (14.3%) has a higher extraction rate than solvent extraction (10.3%) and soxhlet extraction (3%). To explore conditions for the analysis of triacylglycerides (TAG) of mackerel oil by MALDI-TOF MS, using different matrix, laser intensity, sample to matrix ratio and deposited times. The optimum conditions are 2,5-dihydroxybenzoic acid as matrix and laser energy 90%, the sample to matrix ratio (1/1, v/v) and deposited three times, according to peak abundance and profile clarity in the spectrum. MALDI-TOF MS analysis on TAG is matched with the online database LIPID MAPS Online Tools to identify TAG in mackerel oil. The main TAG molecules and the assigned fatty acid compositions in mackerel oil are identified as 25 TAGs: m/z 799.7 (C46:1, MPP1), 801.7 (C46:0, MPP), 813.6 (C48:8), 815.6 (C48:7, LM1D6/M1M1E5), 817.6 (C48:6, LP1E5/LLn3Ln3), 819.6 (C48:5, MME5), 825.7 (C48:2, MPL2/PP1P1), 827.7 (C48:1, PPP1/MPO1), 829.7 (C48:0, PPP), 839.6 (C50:9), 841.6 (C50:8, M1M1D6/LLn3E5), 843.6 (C50:7, LP1D6/M1P1E5), 845.7 (C50:6, MMD6/MP1E5), 849.7 (C50:4, ML2L2), 851.7 (C50:3, PPLn3/PP1L2), 853.7 (C50:2, PPL2/PP1O1), 867.6 (C52:9, LLn3D6/M1Ln3E5), 869.7 (C52:8, M1P1D6/MLn3E5), 875.7 (C52:5, PPE5/PL2Ln3), 877.7 (C52:4, PL2L2), 879.7 (C52:3, PO1L2), 889.6 (C54:12), 891.6 (C54:11, LE5D6/M1E5E5), 913.6 (C54:0), 915.6 (C56:13).When comparing with the TAG of menhaden and cod liver oils, m/z 813.6, 815.6, 839.6, 843.6, 889.6, 891.6 are unique for mackerel oil, and m/z 799.7, 825.7, 827.7, 829.7, 845.7, 849.7, 853.7, 869.7, 875.7, 877.7, 879.7 are co-present in menhaden and cod liver oils. The TAG shared in vegetable oils of soybean oil and sunflower oil are apparently different from that of the mackerel oil . The TAG molecule of mackerel oil have potential power of discrimination for species identification and determination of adulterated products with lower priced vegetable oils in fish oil or fish oil feed. At the same time, the phosphatidylcholine (PC) of mackerel oil was analyzed by MALDI-TOF MS, which were m/z 803.6, 805.6, 831.7, 833.7, and the molecular composition was PC (18:2/20:5), PC (16:0/22:6), PC (18:1/22:6), PC (20:1/22:6).
Appears in Collections:[Department of Food Science] Dissertations and Theses

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