反相高效液相色谱法同时测定胸腔积液中3种氨基酸和3种修饰核苷

发布时间：2018-03-03 17:26

本文选题：反相高效液相色谱法　切入点：氨基酸　出处：《理化检验(化学分册)》2014年01期 　论文类型：期刊论文

【摘要】：提出了反相高效液相色谱法同时测定胸腔积液中3种氨基酸和3种修饰核苷的方法。样品用甲醇提取,离心后上清液以Waters sunfire C18色谱柱为分离柱,不同体积比的甲醇和10mmol·L-1乙酸铵溶液(pH 4.6)的混合溶液为流动相进行梯度洗脱,在278nm处测定酪氨酸、色氨酸,257nm处测定2′-脱氧鸟苷、次黄嘌呤核苷和鸟嘌呤核苷,215nm处测定苯丙氨酸。苯丙氨酸的线性范围为2~2 000μmol·L-1,其余5种化合物为1~1 000μmol·L-1。方法的检出限(3S/N)在0.008 3~0.50μmol·L-1之间。加标回收率在85.8%~106%之间。
[Abstract]:A method for simultaneous determination of three amino acids and three modified nucleosides in pleural effusion by reversed phase high performance liquid chromatography (RP-HPLC) was developed. The samples were extracted by methanol and the supernatants were separated by Waters sunfire C18 column after centrifugation. The mixture of methanol and 10 mmol 路L -1 ammonium acetate solution (pH 4.6) was used as mobile phase for gradient elution. Tyrosine was determined at 278nm, and 2H-deoxyguanosine was determined at 257nm by tryptophan. Phenylalanine was determined at 215 nm of Hypoxanthine and guanosine. The linear range of phenylalanine was 2 000 渭 mol 路L -1 for phenylalanine and 1 000 渭 mol 路L -1 for the other five compounds. The detection limit of the method was between 0.008 3 渭 mol 路L -1 and 0.50 渭 mol 路L -1. The recoveries of phenylalanine were in the range of 85.810 6%.
【作者单位】：浙江工业大学化学工程与材料学院;浙江省人民医院呼吸内科;
【基金】：浙江省自然科学基金资助项目(M203030;Y4110536) 浙江省公益性技术应用研究(分析测试)项目(2011C37040,2011C37009)
【分类号】：R561.3;O657.72

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