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  

Simultaneous Determination of 3 Amino Acids and 3 Modified Nucleosides in Pleural Effusion by RP-HPLC

作　　者：汤晓东[1] 王宏[2] 王丽丽[1] 刘会君[1] 潘再法[1] 

机构地区：[1]浙江工业大学化学工程与材料学院,杭州310014 [2]浙江省人民医院呼吸内科,杭州310014

出　　处：《理化检验（化学分册）》2014年第1期63-66,共4页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

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)

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10 mmol·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278 nm处测定酪氨酸、色氨酸,257 nm处测定2 '-脱氧鸟苷、次黄嘌呤核苷和鸟嘌呤核苷,215 nm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1～1 000 μmol·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.A method of RPHPLC for the simultaneous determination of 3 amino acids and 3 modified nucleosides in pleural effusion was proposed.The sample was extracted with methanol.After centrifugation,the supernatant obtained was use for HPLC analysis using Waters sunfire C18 column as stationary phase,and mixtures of methanol and 10 mmol · L-1 ammonium acetate solution (pH 4.6) in different ratios as mobile phase in the gradient elution.UV-detection at 278 nm was adopted in the determination for tyrosine and tryptophan,257 nm for 2'-deoxyguanosine,inosine and guanosine,and 215 nm for phenylanine.Linearity ranges were found between 2 2 000 μmol · L-1 for phenylanine and 1-1 000 μ mol· L-1 for the other 5 compounds.Values of detection limit (3S/N) of the method found were in the range of 0.008 3-0.50 μ mol · L-1.Values of recovery found by standard addition method were in the range of 85.8％-106％.

关 键 词：反相高效液相色谱法 氨基酸 修饰核苷 胸腔积液 

分 类 号：O657.72[理学—分析化学] O657.32[理学—化学]