- 學位論文
以液相層析串聯質譜法分析臺灣婦女泌乳期間母乳寡醣與三酸甘油酯之變化
Changes of oligosaccharide and triacylglycerol in human milk during lactation in Taiwan using LC-MS/MS
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摘要
母乳是新生兒的唯一天然營養來源，母乳寡醣與脂質是母乳中第三及第二豐富的成分。本研究使用液相層析串聯質譜法解析母乳中21種母乳寡醣以及57個三酸甘油酯組成與脂肪酸分布，將泌乳期前52週分為八期，觀察泌乳期間之變化。樣品來自成大醫院與臺大醫院招募89位母親所提供237個母乳，分別在出生後第 1、2.5、4、5、8、16、24和52週進行收集。母乳寡醣以2’-fucosyllactose (2’-FL) 占19.5%最為豐富，其次為3-fucosyllactose (3-FL) 占15.6%、Lacto-N-fucopentaose II (LNFP II) 占14.0%與Lacto-N-tetraose (LNT) 占12.1%，總和占比超過60%。母乳寡醣總濃度由出生週次10.87 mg/mL下降至泌乳期第24週7.09 mg/mL。母乳中岩藻糖基化母乳寡醣到第52週時顯著增加14.3%，唾液酸化和無岩藻糖基化母乳寡醣則顯著下降5%及9.3%。藉由特徵碎片辨別寡醣上之抗原表位推測母親之基因型，發現樣品主要來自於Se+Le+型的臺灣母親。另外，同時發現母乳中主要三酸甘油酯為β-OPO 與β-OPL在各泌乳期與不同母親間濃度皆維持恆定。母乳中主要的脂肪酸Oleic acid、Palmitic acid及Linoleic acid，以二酸甘油酯特徵碎片離子強度發現近60%的 Palmitic acid鍵結母乳三酸甘油酯sn-2位置，不飽和脂肪酸Oleic acid及Linoleic acid則傾向鍵結在sn-1/3位置。最後，研究中提供可能影響母乳寡醣及三酸甘油酯組成變化的原因，分別討論母親食用營養補充品、年齡、血型、妊娠週數、胎次等影響。對於母乳中可能交互影響的因子尚待確認，建議未來除了收集更長時間之泌乳週數外，對於影響母乳寡醣及三酸甘油酯組成因子應再進行深入研究。本研究結果提供臺灣婦女泌乳期間母乳寡醣及三酸甘油酯成分資料的初探，將有助於臺灣婦女母乳中成分資料的建立。
並列摘要
Human milk is a sole nutrient source for infants. Human milk oligosaccharides (HMO) and fats are composed of a large number of structures that in total are the third and second, respectively, abundant group of compounds in human milk, in addition to lactose and proteins. This study usd a UHPLC-MS/MS platform to reveal the specific ingredients in human milk, including oligosaccharides and triacylglycerols composition. A total of 237 human milk samples were obtained from 89 women who delivered healthy full-term infants by NCKUH and NTUH recruited. The milk sample were collected on postnatal week 1, 2.5, 4, 5, 8,16, 24 and 52. The main HMO are 2’-fucosyllactose (2’-FL), which accounts for 19.5% of the total HMO, followed by 3-fucosyllactose (3-FL, 15.6%), Lacto-N-fucopentaose II (LNFP II, 14.0%) and Lacto-N-tetraose (LNT, 12.1%). They are over 60% of total HMO. The total concentrations of HMO in these samples were found to be 10.87 mg/mL on postnatal week 1, and declined to 7.09 mg/mL on week 24. During the lactation fucosylated HMO increased 14.3% significantly, and both sialylated and nonfucosylated neutral HMO were significant decreased 5% and 9.3%. Identifying epitopes on oligosaccharides from characteristic fragments inferred the Le+Se+ type was the most abundant in Taiwanese mothers. As regards human milk triacylglycerols (TAG), both β-OPO and β-OPL were founding the main triacylglycerols in human milk at different donors and lactation periods. Oleic acid, palmitic acid, and linoleic acid were the main fatty acid of Taiwanese human milk. Over 60% of the palmitic acid was esterified at the TAG stereo-specifically numbered sn-2 position, with preferential positioning of oleic acid and linoleic acid at the TAG sn-1,3 positions. In addition, we investigated mothers’ dietary habits. The results found that mother’s consuming dietary supplements during pregnant or lactation, age, blood type, gestational weeks, and parity may affect the oligosaccharides and triacylglycerols profiles in human milk. Our study revealed the relationship among Taiwanese human milk oligosaccharides, triacylglycerols, and maternal dietary supplementation. It will contribute the understanding of the composition of Taiwanese human milk.
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