色谱

色谱

• 研究论文 • 上一篇    下一篇

纳米纤维固相萃取-高效液相色谱法测定血浆中的5-羟色胺

周小玲1, 王羽1, 陈利琴1, 康学军1,2*   

  1. 1. 东南大学儿童发展与学习科学教育部重点实验室, 江苏 南京 210096; 2. 东南大学苏州研究院 苏州环境与生物安全重点实验室, 江苏 苏州 215123
  • 收稿日期:2011-05-27 修回日期:2011-06-14 出版日期:2011-11-28 发布日期:2011-12-25
  • 通讯作者: 康学军,博士,教授,主要研究方向为生物分析化学.
  • 基金资助:

    国家自然科学基金面上项目(81172720)、苏州市应用基础研究项目(SYJG0912, SYN201006, SG201028)和江苏省工业支撑项目(BE2010088).

Determination of 5-hydroxytryptamine in plasma by nanofiber solid phase extraction-high performance liquid chromatography

ZHOU Xiaoling1, WANG Yu1, CHEN Liqin1, KANG Xuejun1,2*   

  1. 1. Key Laboratory of Child Development and Learning Science (Ministry of Education), Southeast University, Nanjing 210096, China; 2. Research Institute of Southeast University in Suzhou, Suzhou Key Laboratory of Environment and Biosafety, Suzhou 215123, China
  • Received:2011-05-27 Revised:2011-06-14 Online:2011-11-28 Published:2011-12-25

PDF

547

被引次数

19

摘要： 基于纳米纤维的富集作用,建立了血浆中5-羟色胺(5-HT)的柱前衍生高效液相色谱-电化学检测(HPLC-ECD)分析方法。用10%(v/v)高氯酸溶液沉淀血浆蛋白,离心后取上清液,用0.1 mol/L的四苯硼酸钠溶液调节pH值至8.5,加入衍生剂邻苯二甲醛溶液于30 ℃衍生4 min,经纳米纤维固相萃取柱净化富集后,以pH 5.4的0.05 mol/L磷酸二氢钾缓冲液（含0.25 mmol/L乙二胺四乙酸)-甲醇(60:40, v/v)为流动相在C18色谱柱(150 mm×4.6 mm, 5 μm)上进行分离,进样量为20 μL。结果表明,在高、中、低3个加标水平下,血浆中5-HT的加标回收率为95.6%～101.4%,日内与日间相对标准偏差均小于5%(n=3)。方法的定量线性范围为5～500 μg/L,检出限为1 μg/L。该方法操作简单、快速,具有较高的灵敏度和较高的重现性,能够用于血浆中5-HT的含量测定。

关键词: 5-羟色胺, 高效液相色谱, 固相萃取, 纳米纤维, 血浆

Abstract: 1. Key Laboratory of Child Development and Learning Science (Ministry of Education), Southeast University, Nanjing 210096, China; 2. Research Institute of Southeast University in Suzhou, Suzhou Key Laboratory of Environment and Biosafety, Suzhou 215123, China) Abstract: A novel packed-nanofiber solid phase extraction coupled with high performance liquid chromatography-electrochemical detection (HPLC-ECD) method was established for the determination of 5-hydroxytryptamine (5-HT) in plasma. A 10% (v/v) HClO4 solution was used to precipitate the protein in plasma samples. After homogenizing for 1 min and centrifuging for 10 min at 12000 r/min, the supernatant was adjusted to pH 8.5 with 0.1 mol/L sodium tetraphenylborate and then derivatized with o-phthalaldehyde (OPA) solution at 30 ℃ for 4 min. The solution was then purified and preconcentrated by the packed-nanofiber solid phase extraction column, in which methanol was used as the eluent. The analyte was analyzed by HPLC-ECD. The chromatographic separation was achieved on a Shimadzu C18 column with pH 5.4 0.05 mol/L phosphate buffer solution (containing 0.25 mmol/L ethylene diamine tetraacetic acid)-methanol (60:40, v/v) as mobile phase. The linear range was 5~500 μg/L with the correlation coefficient (r2) of 0.9996. The limit of detection (S/N=3) was 1 μg/L. And the spiked recoveries were in the range of 95.6%~101.4% with the relative standard deviations (RSDs) of intra-day and inter-day assays below 5% (n=3). On the basis of the advantages of simplicity, high sensitivity and good reproducibility, this method can be used for the determination of 5-HT in human plasma.

Key words: 5-hydroxytryptamine, high performance liquid chromatography (HPLC), plasma, solid phase extraction, nanofibers