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Application of combination of capillary electrophoresis with laser-induced fluorescence:measurement of glutamate and arginine in PAG microdialytes of conscions morphine-withdrawal rats

Cao Junli, Zhang Yahai, Gu Jun, Zhou Wenhua, Yang Guodong, Zeng Yinming

Department of Anesthesiology，Affiliated Hospital of Xuzhou Medical College，Jiangsu Province Key Laboratory of Anesthesiology. Xuzhou 221002, China；Ningbo Addiction Research and Treatment Center. Ningbo 315010,China

Abstract

Microdialysis technique in free-moving animals can be used to monitor continuously the chan-ges of many extracellular neurotransmitters in certain brain areas and study the relationship between neuro-transmitter and functions. Using detection of capillary electrophoresis combined with laser-induced fluores-cence (CE-LIF) further improves the above-mentioned technique. In the present study, fluorescein isothio-cyanate (FITC ) was used to derivatizate amino acid in very low concentration. We found that increasing de-rivatization temperature could shorten derivatization time and that the derivatizative efficiency was not differ-ent from that when experiment was performed under the traditional derivatization condition (room tempera-ture for 16 h). We also got an optimized condition of amino acid derivatization with FITC at 30℃ waterbath for 5 h. Using the optimized condition of amino acid derivatization, we investigated the changes in L- arginine (L-Arg) and L-glutamate (L-Glu) concentration in periaqueductal gradually. These changes were in correspondence with the scores of morphine withdrawal symptom.

Key words: Capillary electrophoresis;Microdialysis glutamate (Glu);Arginine (Arg); Periaqueductal gray matter ;

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Citing This Article：

Cao Junli, Zhang Yahai, Gu Jun, Zhou Wenhua, Yang Guodong, Zeng Yinming. Application of combination of capillary electrophoresis with laser-induced fluorescence:measurement of glutamate and arginine in PAG microdialytes of conscions morphine-withdrawal rats. Acta Physiol Sin 2003; 55 (5): (in Chinese with English abstract).