Simultaneous Determination of 3 Amino Acids and 3 Modified Nucleosides in Pleural Effusion by RP-HPLC
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2~2 000 μmol·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 3~0.50 μmol·L-1之间。加标回收率在85.8%~106%之间。
Abstract
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 μmol·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 μmol·L-1. Values of recovery found by standard addition method were in the range of 85.8%-106%.
中图分类号 O652.63
所属栏目 工作简报
基金项目 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)
收稿日期 2013/1/30
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备注汤晓东(1988-),男,浙江武义人,硕士研究生,研究方向为色谱分析。
引用该论文: TANG Xiao-dong,WANG Hong,WANG Li-li,LIU Hui-jun,PAN Zai-fa. Simultaneous Determination of 3 Amino Acids and 3 Modified Nucleosides in Pleural Effusion by RP-HPLC[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2014, 50(1): 63~66
汤晓东,王宏,王丽丽,刘会君,潘再法. 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[J]. 理化检验-化学分册, 2014, 50(1): 63~66
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【2】张春晓,叶庆林,蒋滢.肺癌患者尿液中13种游离氨基酸分析[J].福建医科大学学报, 2002,36(4):444-445.
【3】PROENZA A M, OLIVER J, PALOU A, et al. Breast and lung cancer are associated with a decrease in blood cell amino acid content[J]. The Journal of Nutritional Biochemistry, 2003,14(3):133-138.
【4】SCHWARZ E L, ROBERTS W L, PASQUALI M. Analysis of plasma amino acids by HPLC with photodiode array and fluorescence detection[J]. Clinica Chimica Acta, 2005,354(1):83-90.
【5】杨进,陆友金,檀华容.肺癌患者与正常人血、尿中氨基酸谱的对比研究[J].临床肺科杂志, 2007,12(5):429-430.
【6】MAEDA J, HIGASHIYAMA M, IMAIZUMI A, et al. Possibility of multivariate function composed of plasma amino acid profiles as a novel screening index for non-small cell lung cancer: a case control study[J]. Bmc Cancer, 2010,10(1):690-697.
【7】LIMBACH P A, CRAIN P F, MCCLOSKEY J A. Summary: the modified nucleosides of RNA[J]. Nucleic Acids Research, 1994,22(12):2183-2196.
【8】陈英杰,郑育芳,王凝芳,等.尿中核苷检测在胃癌诊断中的意义[J].癌症, 2003,22(5):537-540.
【9】MCENTIRE J E, KUO K C, SMITH M E, et al. Classification of lung cancer patients and controls by chromatography of modified nucleosides in serum[J]. Cancer Research, 1989,49(4):1057-1062.
【10】XU Guo-wang, STEFANO D C, LIEBICH H M, et al. Reversed-phase high-performance liquid chromatographic investigation of urinary normal and modified nucleosides of cancer patients[J]. Journal of Chromatography B: Biomedical Sciences and Applications, 1999,732(2):307-313.
【11】吴逸明,周舫,吴拥军,等.高效液相色谱法测定肺癌患者尿中假尿嘧啶核苷和肌苷量比值[J].郑州大学学报:医学版, 2002,37(4):433-435.
【12】ZANINI D, SCHMATZ R, PIMENTEL V C, et al. Lung cancer alters the hydrolysis of nucleotides and nucleosides in platelets[J]. Biomedicine & Pharmacotherapy, 2012,66:40-45.
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