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Chinese Journal of Chromatography
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Chinese Journal of Chromatography  2015, Vol. 33 Issue (9): 981-987    DOI: 10.3724/SP.J.1123.2015.04049
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Determination of L-dopa and dopamine in rat brain microdialysate by ultra high performance liquid chromatography-tandem mass spectrometry using stable isotope-coded derivatization coupled with dispersive liquid-liquid microextraction
QI Weimei1, ZHAO Xian-en2, QI Yong1, SUN Zhiwei2, CHEN Guang2, YOU Jinmao2,3, SUO Yourui3
1. Chemical Department, Laiwu Vocational and Technical College, Laiwu 271100, China;
2. Shandong Provincial Key Laboratory of Life-organic Analysis, Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, China;
3. Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China
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Abstract 

The sensitive detection method of levodopa (L-DOPA) and dopamine (DA) in rat brain microdialysate of Parkinson's disease (PD) is an essential tool for the clinical study and attenuated synergistic drug screening for L-DOPA from traditional Chinese medicines. Using d0/d3-10-methyl-acridone-2-sulfonyl chloride (d0/d3-MASC) as stable isotope derivatization reagent, a novel ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for L-DOPA and DA by stable isotope-coded derivatization coupled with ultrasonic-assisted dispersive liquid-liquid microextraction (UA-DLLME). d0-MASC (light) and d3-MASC (heavy) were used as derivatization reagents for microdialysate samples and standards, respectively. Mixtures of the two solutions were prepared by UA-DLLME for UHPLC-MS/MS analysis with multiple reaction monitoring (MRM) mode. With d3-MASC heavy derivatives as internal standards for corresponding light derivatives from samples, the stable isotope internal standard quantification for L-DOPA and DA was carried out. The stable derivatives were obtained in aqueous acetonitrile (pH 10.8 sodium carbonate-sodium bicarbonate buffer) at 37 ℃ for 3.0 min, and then were separated within 2.0 min using gradient elution. Linear range was 0.20-1500.0 nmol/L (R> 0.994). LODs were 0.005 and 0.009 nmol/L for DA and L-DOPA (S/N=3), respectively. This method was validated, and it showed obvious advantages in comparing with the reported methods in terms of sensitivity, analysis speed and anti-matrix interference. This method has been successfully applied to the study of effect of Shouwu Fang on L-DOPA and DA concentration fluctuations in PD rat brain microdialysate.

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Articles by authors
QI Weimei
ZHAO Xian-en
QI Yong
SUN Zhiwei
CHEN Guang
YOU Jinmao
SUO Yourui
Key wordsrat in vivo microdialysis   mass spectrometry sensitization analysis   ultra high performance liquid chromatography (UHPLC)   Showwu Fang   drug screening   Parkinson's disease (PD)     
Received: 2015-04-28;
Cite this article:   
QI Weimei,ZHAO Xian-en,QI Yong et al. Determination of L-dopa and dopamine in rat brain microdialysate by ultra high performance liquid chromatography-tandem mass spectrometry using stable isotope-coded derivatization coupled with dispersive liquid-liquid microextraction[J]. Chinese Journal of Chromatography, 2015, 33(9): 981-987.
 
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