Evaluation of histidine-containing dipeptides in twelve marine organisms and four land animal meats by hydrophilic interaction liquid chromatography with ultraviolet detection
Evaluation of histidine-containing dipeptides in twelve marine organisms and four land animal meats by hydrophilic interaction liquid chromatography with ultraviolet detection
复制标题
采用亲水相互作用液相色谱-紫外检测法评价 12 种海洋生物和 4 种陆地动物肉中的含组氨酸二肽
DOI：
10.1080/10826076.2018.1526803
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发表时间：
2018
影响因子：
1.3
通讯作者：
Tanaka Ryusuke
中科院分区：
文献类型：
--
作者：
Mori Akihiro;Hikihara Risako;Ishimaru Mami;Hatate Hideo;Tanaka Ryusuke
含组氨酸二肽（HCDP）广泛分布于脊椎动物的骨骼肌中，包括海洋和陆地生物。在本研究中，我们开发了一种简单、快速、准确的方法，通过等度洗脱的高效液相色谱 (HPLC) 来鉴定 HCDP。该方法基于亲水相互作用色谱（HILIC），使用磷酸胆碱HILIC柱（250mm × 4.6mm，5μm颗粒），以15mmol/L乙酸铵（pH 4.25）：乙腈：甲醇（体积比3.5：5.85：0.65）为流动相，流速1.0mL/min，以及214nm 的紫外检测。结果，HCDP，如肌肽、鲸氨酸和鹅氨酸，在 20 分钟内分离出来，结果具有足够的重现性和定量性。使用这种方法，检测限 (LOD) 优于以前的方法。肌肽、鲸氨酸和鹅氨酸的 LOD 分别为 0.062、0.085 和 0.100μg/mL。肌肽、鲸氨酸和鹅氨酸的线性范围为 0.207–500、0.283–500 和 0.334–500μg/mL。平均回收率分别为 99.9%、91.7% 和 100.1%。我们的方法用于评估海洋和陆地生物中的 HCDP。大型中上层洄游鱼类含有大量鹅氨酸。与鱼类相比，陆地和水生哺乳动物都含有大量肌肽。对小须鲸、日本鳗鱼、猪和鸡中的 Balenine 进行了定量。
Histidine-containing dipeptides (HCDP) are widely distributed in the skeletal muscle of vertebrates, including marine and terrestrial organisms. In this study, we developed a simple, rapid, and accurate method for identifying HCDP by high performance liquid chromatography (HPLC) with isocratic elution. The method based on hydrophilic interaction chromatography (HILIC) using phosphorylcholine HILIC column (250 mm × 4.6 mm, 5 µm particles), 15 mmol/L ammonium acetate (pH 4.25):acetonitrile:methanol (3.5:5.85:0.65 by volume) as the mobile phase at 1.0 mL/min, and ultraviolet detection at 214 nm. As a result, HCDP, such as carnosine, balenine, and anserine, were separated in 20 min and the results were sufficiently reproducible and quantitative. Using this method, the limit of detection (LOD) was superior to previous methods. The LOD of carnosine, balenine, and anserine were 0.062, 0.085, and 0.100 µg/mL. The linearity range of carnosine, balenine, and anserine were 0.207–500, 0.283–500, and 0.334–500 µg/mL. The average recovery of that were 99.9, 91.7, and 100.1%. Our method was used to evaluate HCDP in marine and terrestrial organisms. Large pelagic migratory fish contained large amounts of anserine. Both land and aquatic mammals contained large amounts of carnosine compared with fish. Balenine was quantitated in minke whales, Japanese eels, pigs, and chickens.