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高效液相色谱法测定人工脑脊液中谷氨酸的浓度
  • ISSN号:1001-6821
  • 期刊名称:《中国临床药理学杂志》
  • 时间:0
  • 分类:R965.2[医药卫生—药理学;医药卫生—药学]
  • 作者机构:[1]沈阳药科大学生命科学与生物制药学院,沈阳110016, [2]卫生部北京医院药学部,北京100730
  • 相关基金:药物使用安全与输血安全相关技术与标准研究基金资助项目(200902008)
中文摘要:

目的建立高效液相色谱测定大鼠脑脊液中谷氨酸含量的方法。方法微透析实验收集到的脑脊液样品用邻苯二甲醛、β巯基乙醇衍生化反应后,高效液相色谱法分离。用RP18色谱柱,流动相A为50 mmol.L-1乙酸钠(乙酸调pH为6.7),流动相B为甲醇/四氢呋喃(97.5/2.5,v/v),用二元梯度洗脱,梯度程序为(T,B%)(0,30)(7,45)(14,30)(20,30),流速1.0 mL.min-1,荧光检测器定量,激发波长340 nm,发射波长450 nm。用高钾脑脊液灌流,考察大鼠纹状体内谷氨酸的变化。结果谷氨酸的线性范围为3.12×10-2~1.00 mg.L-1(γ=0.9998),日内、日间精密度均〈15%,低、中、高浓度回收率分别为(96.6±6.1)%,(99.1±7.7)%,(96.2±4.1)%。高钾人工脑脊液灌流后,谷氨酸明显升高后降低,随后又有升高的趋势。结论本方法准确,稳定,灵敏,适用于微透析样品中谷氨酸的定量分析。

英文摘要:

Objective To develop a sensitive, accurate and specific high performance liquid chromatography (HPLC) method to measure concentration of glutamic acid in rat cerebrospinal fluid. Methods Microdialysis samples were derivatized by o - phthaldialdehyde (OPA) and β - mercaptoethanol, separated by HPLC and quatitated by fluorescence detector. Mobile phase A was 0. 05 mol · L- 1 sodium acetate ( pH adjusted to 6.7 with acetic acid), mobile phase B was methanol/tetrahydrofuran (97.5/2. 5 ,v/v). Gradient program was (T,B%) (0,30) (7,45) ( 14, 30) (20,30). The total flow rate was 1.0 mL · min-1. Excitation wave was 340 nm, and emission wave was 450 nm. Investigating changes of glutamic acid in rat's striatum was used high potassium artificial cerebrospinal fluid (aCSF) perfusion through microdialysis probe. Results The retention time of glutamic acid was 2. 8 rain. Good linearity (3/= 0. 9998) was obtained between 3.12 × 10-2 and 1. O0 mg · L-1. RSD of intra- day and inter- day validation was lower than 15%. Recovery results of QCwere (96.6±6.1)%,(99.1±7-7)%,(96.2±4.1)%, respectively. After perfusion of high potassium aCSF, glutamic acid increased significantly and then decreased to baseline, and there was a trend of increasing after that. Conclusion The method is sensitive, accurate and specific, and can be applied to the quantification of glutamic acid in microdialysis samples of rat.

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期刊信息
  • 《中国临床药理学杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国药学会
  • 主编:韩启德
  • 地址:北京市海淀区学院路38号
  • 邮编:100191
  • 邮箱:cjcp1985@163.com
  • 电话:010-82802540
  • 国际标准刊号:ISSN:1001-6821
  • 国内统一刊号:ISSN:11-2220/R
  • 邮发代号:82-142
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),美国剑桥科学文摘,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:22227