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采用液相色谱/同位素比率质谱法测定未衍生化氨基酸的δ(13)C用于营养研究:日粮非必需氨基酸组成对鱼类个体氨基酸同位素特征的影响

Determination of underivatized amino acid delta(13)C by liquid chromatography/isotope ratio mass spectrometry for nutritional studies: the effect of dietary non-essential amino acid profile on the isotopic signature of individual amino acids in fish.

作者信息

McCullagh James, Gaye-Siessegger Julia, Focken Ulfert

机构信息

Chemistry Research Laboratory, Oxford University, Mansfield Road, Oxford OX1 3TA, UK.

出版信息

Rapid Commun Mass Spectrom. 2008 Jun;22(12):1817-22. doi: 10.1002/rcm.3554.

Abstract

This study provides data for the effect of dietary non-essential amino acid composition on the delta(13)C values of individual amino acids in rainbow trout (Oncorhynchus mykiss) using liquid chromatography coupled to isotope ratio mass spectrometry (LC/IRMS). In this experiment, trout were reared either on a control diet or on three experimental diets, differing in the composition of non-essential/conditionally essential amino acids, for a period of 6 weeks. The control diet was a commercial trout starter feed with fish meal as the main protein source. The experimental diets contained no protein, only synthetic amino acids. Diet 1 resembled the composition of fish meal in both essential and non-essential amino acids, Diet 2 had all essential amino acids, but cysteine, glycine, proline and tyrosine were replaced by the corresponding amounts of their precursors, and in Diet 3 all non-essential amino acids were replaced by glutamate. LC/IRMS was used for the determination of delta(13)C values of individual amino acids from diets and tissues without derivatization. Diet affected the delta(13)C of individual amino acids in fish. For fish on Diets 1-3 amino acid delta(13)C values showed a similar trend: phenylalanine showed very little change from diet to body tissue. Arginine, lysine, tyrosine and proline showed strong depletion from diet to body tissue and glycine, alanine, aspartate and serine all showed variable but strong enrichment in (13)C. Improvements are necessary before all amino acid delta(13)C values can be determined; however, this study demonstrates that measuring amino acid isotopic signatures by LC/IRMS is a promising new technique for nutritional physiologists.

摘要

本研究利用液相色谱-同位素比率质谱联用仪(LC/IRMS),提供了关于日粮非必需氨基酸组成对虹鳟(Oncorhynchus mykiss)个体氨基酸δ(13)C值影响的数据。在本实验中,虹鳟分别投喂对照日粮或三种实验日粮,为期6周,这三种实验日粮的非必需/条件性必需氨基酸组成有所不同。对照日粮是一种以鱼粉为主要蛋白质来源的商业虹鳟开口饲料。实验日粮不含蛋白质,仅含合成氨基酸。日粮1的必需和非必需氨基酸组成与鱼粉相似,日粮2含有所有必需氨基酸,但半胱氨酸、甘氨酸、脯氨酸和酪氨酸被相应量的前体所替代,日粮3中所有非必需氨基酸均被谷氨酸替代。LC/IRMS用于测定日粮和组织中未经衍生化的个体氨基酸的δ(13)C值。日粮影响鱼体中个体氨基酸的δ(13)C值。对于投喂日粮1-3的鱼,氨基酸δ(13)C值呈现相似趋势:从日粮到鱼体组织,苯丙氨酸变化很小。精氨酸、赖氨酸、酪氨酸和脯氨酸从日粮到鱼体组织呈现强烈亏损,而甘氨酸、丙氨酸、天冬氨酸和丝氨酸均呈现不同程度但强烈的(13)C富集。在能够测定所有氨基酸的δ(13)C值之前仍需改进;然而,本研究表明,通过LC/IRMS测量氨基酸同位素特征对营养生理学家来说是一项有前景的新技术。

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