柱前衍生化高效液相色谱法分析大鼠体内D-丝氨酸和L-丝氨酸  

Determination of D-serine and L-serine in Rats by High Performance Liquid Chromatography with Pre-Column Derivatization

作　　者：余海立 张彦 严飞 冯雪萍 李春阳 张双庆 YU Hai-li;ZHANG Yan;YAN Fei;FENG Xue-ping;LI Chun-yang;ZHANG Shuan-qing(The Hubei Provincial Key Laboratory of Yeast Function,Yichang 443003,China;National Institute for Nutrition and Health,Chinese Center for Disease Control and Prevention,Beijing 100050,China)

机构地区：[1]酵母功能湖北省重点实验室,湖北宜昌443003 [2]中国疾病预防控制中心营养与健康所,北京100050

出　　处：《中国食物与营养》2022年第9期57-60,共4页Food and Nutrition in China

基　　金：湖北省自然科学基金项目(项目编号:2018CFB612)。

摘　　要：目的:建立柱前衍生化高效液相色谱紫外检测法分析大鼠血浆和脑组织中D-丝氨酸和L-丝氨酸的方法,比较对照组与缺硒组大鼠血浆和脑组织中D-丝氨酸和L-丝氨酸含量的差异,研究D-丝氨酸和L-丝氨酸与硒缺乏的关系。方法:衍生化试剂:邻苯二甲醛和N-乙酰-L-半胱氨酸,衍生温度为室温,反应时间为5 min。采用Waters X bridge Amide色谱柱(250 mm×4.6 mm,3.5μm),进行梯度洗脱,流速1.0 mL/min、柱温30℃、检测波长340 nm。结果:D-丝氨酸和L-丝氨酸分别在2~14μg/mL和8~56μg/mL范围内线性关系良好,相关系数(R^(2))分别为0.9996、0.9994,加样回收率为97.29%~101.88%。结论:该方法快速灵敏,可用于大鼠体内D-丝氨酸和L-丝氨酸含量的测定。对照组和缺硒组大鼠血浆中L-丝氨酸含量有显著性差异。【Objective】 To compare the serine levels between normal and selenium-deficient rats. 【Method】 A performance liquid chromatography(HPLC) with a ultraviolet detector method for the determination ofD-serine andL-serine in serum and brain tissues of rats was established. The method involved the pre-column derivatization of amino acids using O-phthalaldehyde and N-acetyl-L-cysteine for 5min at room temperature. HPLC analysis conditions were Waters X bridge Amide column(250 mm × 4. 6 mm, 3. 5 μm), flow rate of 1. 0mL/min, column temperature of 30℃, detection wavelength of 340 nm and mobile phase of ammonium acetate(50 mmol/L) and acetonitrile. 【Result】 The results showed that bothD-serine andL-serine showed good linearity in the range of 2-14 μg/mL and 8-56 μg/mL with correlation coefficient values of 0. 999 6 and 0. 999 4, respectively. The recovery was 97. 29%-101. 88%. The method was rapid,sensitive and can be applied to the determination of the content ofD-serine andL-serine in rats. 【Conclusion】 Levels of plasmaL-serine had significant differences between normal and selenium-deficient rats.

关 键 词：D-丝氨酸 L-丝氨酸 高效液相色谱法 硒 

分 类 号：O657.72[理学—分析化学] R446[理学—化学]