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1、 ? 2013 ? 9 ? 30 ? 9 ? Chin JMAP, 2013 September, Vol.30 No.9 ?929? ? ? ? pH ? ?1?2*?1?1?1?1(1.? ? 415000?2.? 510520) ? ? 5 ?-?(HPLC-FD)?(TMPP)? pH? ? Agilent 1100 ?ZORBAX SB-Aq Cl8? 357 nm? 455 nm? 50 mmoLL1?(pH=6.5)? 1 mLmin1?12 ? 6 ?HPLC-FD ? 5 ? ? TMPP ? pH ?(10 mgkg1)? Glu ? Asp ? Tau ? GABA ?
2、TMPP ? ?-? pH ? ?R969.11 ?A ?1007-7693(2013)09-0929-05 Influence of Nasal Tetramethylpyrazine Phosphate pH-Sensitive in Situ Gel on Amino Acid Content in Rats Brain of Acute Cerebral Ischemia Model LIU Hongwei1, YAN Yilin2*, SHE Jinming1, REN Minqiong1, TAN Min1, CHEN Jing1(1.Changde Vocational Tech
3、nical College, Changde 415000, China; 2.Guangdong Food and Drug Vocational College, Guangzhou 510520, China) ABSTRACT: OBJECTIVE To establish a determination method for 5 kinds of amino acid in rats with high performance liquid-fluorescence detector. To study the effects of tetramethylpyrazine phosp
4、hate(TMPP) on amino acid content in acute cerebral ischemia rats model. METHODS Microdialysis fluid was determined with Agilent 1100 liquid system fluorescence detector, ZORBAX SB-Aq Cl8 chromatographic column was applied. Excitation wavelength and emission wavelength were set at 357 nm and 455 nm,
5、respectively, mobile phase consisted of methanol and 50 mmolL1 sodium acetate buffer (pH=6.5), with gradient elution, the flow rate was 1 mLmin1. Tweleve male rats randomly divided into model control group and treatment group, each group had 6. Microdialysis was used to collect samples in striatum o
6、f rats area after dialysate, five kinds of amino acid concentration was determined by HPLC-FD. RESULTS Glu and Asp levels in TMPP group were decreased significantly compare with control group, however, the amount of Tau and GABA increased. CONCLUSION TMPP can obviously reduce excitatory amino acid c
7、ontent and increase brain inhibitory amino acid content in model rats, which has protect effect on acute ceretral ischemia rats. KEY WORDS: HPLC-FD; tetramethylpyrazine phosphate pH-sensitive in situ gel; amino acid; brain microdialysis; acute cerebral ischemia rat model ?(30801551)?(845105200500118
8、6) ? Tel: 15080603524 E-mail: *? Tel: (020)28854912 E-mail: yilin_ ? ? ? ? ?1?(tetramethylpyrazine?TMP)? ? ?2? (tetramethylpyrazine phosphate?TMPP)? ?930? Chin JMAP, 2013 September, Vol.30 No.9 ? 2013 ? 9 ? 30 ? 9 ? ?3?TMP ?4? ? ? ? ?HPLC-FD? ? ? ? ? ? TMPP ? ? pH ? ? ? ? ? ? ? (10 mgkg1)5?TMPP? ?T
9、MPP? ? 1 ? 1.1 ? Agilent 1100 ?(Agilent ?)?68001 ?(? 1.2 mm?)?BAS ?MD0100 ?MD1001 ?MD1002 ?MD1000 ?(BAS ?)?CMA/12?4 mm membrane window?cut-off 20k dalton(? CMA ?)? 1.2 ? ?(Glu)? ?(Asp)? ?(Gly)?-?(GABA)?(Tau)(Sigma ?)? 2,3,5-?(TTC?)?(?)?(?)? ?(OPA?)?-?(Sigma ?)? ?(? ?20080915)? 1.3 ? ? SD ? 300350 g?
10、SKXK(?) 2008-0002? 2 ? 2.1 ? ? CMA ? 147 mmolL1? 1.2 mmolL1? 2.7 mmolL1? 0.85 mmolL1? 0.22 m ? 2.2 TMPP ? pH ? ? Carbopol ? HPMC ? 24 h ? pH ? 5.5 ? 2.3 ? 2.3.1 ? ? OPA 10 mg ?0.25 mL ? 20 L -?2 mL 0.2 molL1?(pH=9.5)? ? ? 24 h ? 10 L -? 2.3.2 ? ? 5 L ? 15 L? 2 min ? 2.4 ? Agilent ZORBAX SB-Aq Cl8(4.
11、6 mm?250 mm?5 m)? 357 nm? 455 nm? 25 ? ? A ? B ? 50 mmolL1?(pH=6.5)?03 min?25%? A?75%? B?8 min?35%?A? 65%? B? 2325 min? 45%? A? 55%? B?30 min?25%? A?75%? B? 1 mLmin1? 1? 1 ? ? 1 ? A?(Asp? tR=3.017 min? Glu? tR=3.592 min? Gly? tR=10.592 min?Tau?tR=12.701 min?GABA?tR=15.243 min)?B? Fig 1 HPLC spectrum of amino acid Aamino acid standard(Asp: tR=3.017 min; Glu: tR=3.592 min; Gly: tR=10.592 min; T
