Development of a Method for Measuring Phytanic Acid as a Lifestyle-related Disease Biomarker in Rat Serum Using Ultra-fast Liquid Chromatography-Ultraviolet Spectrophotometry Combined with a Modified 2-Nitrophenylhydrazine Derivatization Method
Development of a Method for Measuring Phytanic Acid as a Lifestyle-related Disease Biomarker in Rat Serum Using Ultra-fast Liquid Chromatography-Ultraviolet Spectrophotometry Combined with a Modified 2-Nitrophenylhydrazine Derivatization Method
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DOI：
10.2116/analsci.33.365
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发表时间：
2017-03
影响因子：
1.6
通讯作者：
Aoi Miyamoto;Takahiko Aoyama;M. Okamura;N. Fukuda;T. Ueno;M. Abe;Y. Matsumoto
中科院分区：
文献类型：
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作者：
Aoi Miyamoto;Takahiko Aoyama;M. Okamura;N. Fukuda;T. Ueno;M. Abe;Y. Matsumoto
建立了一种简单快速的超快速液相色谱-紫外分光光度法（UFLC-UV）结合修饰的2-硝基苯肼（2-NPH）衍生化测定大鼠血清中植烷酸（Phy）的方法。血清 Phy 和十七烷酸（内标）在环境温度下用 2-NPH 衍生化 20 分钟，并用正己烷萃取。从反应混合物中提取衍生化 Phy (D-Phy) 和衍生化 IS 后，使用 YMC-Pack C8 柱（150 × 3.0 mm i.d., S-3 μm）使用由乙腈:H_2O (90:10; pH 4.4) 组成的等度流动相以 0.5 mL/min 分离提取物。检测波长为228 nm。在 1–20 μg/mL 范围内观察到线性 (r = 0.9997)。 D-Phy 测量的日内和日间重现性≤13.0%。据我们所知，这是使用 2-NPH 衍生化和 UFLC-UV 定量和定性测量血清 Phy 的第一份报告。该方法可以在温和条件下快速进行。
A simple and rapid ultra-fast liquid chromatography–ultraviolet spectrophotometry (UFLC-UV) method combined with modified 2-nitrophenylhydrazine (2-NPH) derivatization was developed for determining phytanic acid (Phy) in rat serum. Serum Phy and heptadecanoic acid (the internal standard) were derivatized by 2-NPH at ambient temperature for 20 min and extracted in n -hexane. After extracting derivatized Phy (D-Phy) and derivatized IS from the reaction mixture, the extracts were separated with a YMC-Pack C8 column (150 × 3.0 mm i.d., S-3 μm) using an isocratic mobile phase comprised of acetonitrile:H_2O (90:10; pH 4.4) at 0.5 mL/min. The detection wavelength was 228 nm. Linearity was observed over 1–20 μg/mL ( r = 0.9997). The intra- and inter-day reproducibilities of D-Phy measurements were ≤13.0%. To our knowledge, this is the first report of the quantitative and qualitative measurement of serum Phy using 2-NPH derivatization and UFLC-UV. This method can be performed rapidly under mild conditions.