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Chiral is one of nature the essential attribute of one of the. Many with life activities closely related to amino acids, peptides, proteins and so has biological active substances both has one or more chiral center. Because of symmetry and optical active of the of different, of different enantiomers in the life vivo exhibit different physiological activity, their in vivo changes in the distribution and metabolic associated with certain diseases the Formation closely related. Therefore, the of enantiomers separation and determination of, in the life sciences and other relevant areas all have important to significance. This paper, adopt pre-column derivatization reversephase high performance liquid chromatography (HPLC) analysis techniques, right aspartic acid (Asp), glutamic acid (Glu), serine (Ser), octopus amine (OA) enantiomer conducted a hand sexual Split studies, the establishment a biological samples of D-amino acids and (R)-OA method for measuring, and has been applied mouse brain tissue of D-amino acid and the bees brain tissue (R)-OA the determination of, while the are several D-amino acid associated with Parkinson disease-related Xing conducted a preliminary discussed. Dissertation research work mainly includes the following three aspects: 1. Established for the determination biological samples of Trace D-Asp, D-Glu's HPLC method. In order to o-benzene two formaldehyde-N-isobutyryl acyl-L-cysteine ??(OPA-IBLC) pre-column derivatization. Sodium acetate - methanol - acetonitrile mixed solution as mobile phase, the use of gradient elution technology, 2 kinds of amino acids pairs enantiomers within 40 min completely separated. In fluorescence detection, D-Asp, D-Glu the concentration 10 ~ 1000 nmol / L when its concentration and peak area showed a good linear relationship, when SNR is is 3 (S / N = 3) when, D-Asp and D-Glu's detection limits were 2.0 nmol / L, 1.0 nmol / L. For the first time pairs of Parkinson disease (PD) model mice midbrain D-Asp and D-Glu content change conducted a measured, the results found that control group with the model group the midbrain D-Asp content in was no significant Xing difference in, while the D-Glu content showed significant difference. Speculated that in the medium brain tissue li, of endogenous's D-Asp the synthesis of and transhipment not subject to nerve neurotoxin 1 - methyl-4 - phenyl-1 ,2,3,6 - tetrahydro-pyridine (MPTP)'s poisoned still maintained the original dynamic equilibrium, while the D-Glu might possess with the L-type pairs enantiomer similar to the the excitement toxicity of, participate in the pathogenesis of PD. 2. Using o benzene two formaldehyde-N-isobutyryl acyl-D-cysteine ??(OPA-IBDC) for the hand Xing derivatization reagent, methanol - acetonitrile - sodium acetate mixed solution as mobile phase, on a C18 column split up DL-Ser enantiomer. , Respectively, effects of the mobile phase composition, flow rate and column temperature, etc. on the separation the impact of, established the best separation conditions thereof, its separation degree of reach 2.74. D-Ser concentration of In the 30 ~ 6000 nmol / L range, with reference peak area showed a good linear relationship. Will method was applied PD model mice midbrain AND CEREBRAL CORTEX D-Ser content in's measured, the results indicate that, the model group brain tissue middle D-Ser's content in with the control group compared to the overall showed a a clear downward trend, speculated that D-Ser may be involved in of the PD of the pathological physiological processes. 3. Was established one suitable bees brain tissue (R)-OA Method for determination of. In order to o-benzene two formaldehyde-N-acetyl yl - cysteine ??(OPA-NAC) for the hand Xing derivatization reagent, the use of HPLC technology, right OA enantiomer derivatives chiral separation carried out a detailed studied. Optimized the of derivative conditions and the separation condition, studied of the medium pH, derivative reagent dosage, reaction time, mobile phase composition, column temperature, flow rate and other on the separation selectivity of. In the established best separation conditions under, OA enantiomers Derivatives on within 30min to get better separation, separation degree of of 2.28. Will method was applied honeybee brain tissue middle (R)-OA Determination of content of, high sensitivity, reproducibility Xing good, the operation simple and convenient, results are reliable.
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