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目的:建立测定多巴胺在人工脑脊液中含量的测定方法,考察多巴胺在加入稳定剂后人工脑脊液中的稳定性。方法:采用HPLC-电化学检测法,考察多巴胺在加入稳定剂后于冷藏、冷冻及反复冻融不同条件下的稳定性。结果:多巴胺的保留时间为6.2min,线性范围是0.05~5nmol·L~(-1),日内与日间精密度均<5%,回收率在99.9%~101.6%之间。加入稳定剂后,多巴胺人工脑脊液溶液稳定性显著提高,4℃冰箱内至少能稳定放置3d,在-70℃冰冻条件下至少能稳定放置1周,样品反复冻融3次稳定。大鼠给予氟哌啶醇后,前额叶皮层中多巴胺含量基本无影响,伏隔核中多巴胺显著升高。结论:加入稳定剂后,多巴胺的稳定性显著提高,适用于脑微透析样品中多巴胺的定量分析。
OBJECTIVE: To establish a method for the determination of dopamine in artificial cerebrospinal fluid (CSF), and investigate the stability of dopamine in artificial cerebrospinal fluid (CSF) after adding stabilizers. Methods: The stability of dopamine under different conditions of freezing, freezing and repeated freezing and thawing was investigated by HPLC-electrochemical detection. Results: The retention time of dopamine was 6.2 min and the linear range was 0.05 ~ 5 nmol·L -1. The intra- and inter-day precision was less than 5% and the recovery was between 99.9% and 101.6%. Adding stabilizers, the stability of dopamine artificial cerebrospinal fluid solution significantly increased, at least 4 ℃ refrigerator can be placed in stable at least 3d, frozen at -70 ℃ for at least 1 week can be stably placed, frozen and thawed repeatedly three times the sample stability. After administration of haloperidol in rats, dopamine levels in the prefrontal cortex had almost no effect, and dopamine in the nucleus accumbens increased significantly. CONCLUSION: The stability of dopamine increased significantly with the addition of stabilizer, which is suitable for the quantitative analysis of dopamine in brain microdialysis samples.