Determination Of Nervus Excitability Amino Acids In Rats By HPLC

OBJECT:For comparing the amino acids levels between normal and depressedand evaluating their relationships with the severity of depression, weestablished a HPLC method for determination of D.L-Asp, Glu and Serin serum and brain tissues of rats.METHODS:Pro-column derivated with O-phthalaldehyde (OPA) andN-acetyl-L-cysteine (NAC), the samples were analyzed by a PhenomenexC18column (250mm×4.6mm,5μm).50mmol/L ammoniumacetate-methanol (96:4,v/v) as mobile phase isocratic elution, fluorescencedetected at Ex:350nm, Em:450nm.RESULTS:The resolution of the three enantiomers were1.21,2.20and1.53,respectively. D-amino acids had good linear relationships in the range of5~1000ng/mL, the correlation coefficients were higher than0.9997;L-amino acids had good linear relationships in the range of25~5000ng/mL, the correlation coefficients were higher than0.9998. CONCLUSION:We have established a successfull method by the isocratic elutionRP-HPLC associated with O-phthalaldehyde and N-acetyl-L-cysteine todetect D.L-Asp, Glu and Ser in serum and brain tissues of rats, to comparethe amino acids levels between normal and depressed and to evaluatedtheir relationships with the severity of depression. Found that in the serumand hypothalamus was reduced,L-ser content was obviously descentedwhile D-ser content was obviously rised,and it was significant genderdifference(P<0.05). However, there had no obvious changes of Glu andAsp. In the serum, the L-ser content was obviously descented,or D-sercontent was obviously rised could provide effective evidence for humandepression disease research and clinical diagnosis of disease.