成年大鼠脑透析液和血清中褪黑素含量的测定
发布时间：2018-10-18 13:26
【摘要】：目的采用高效液相色谱(HPLC)-荧光法和酶联免疫(ELISA)法测定成年大鼠脑微透析液及血清中的褪黑素(MLT)含量,比较2种检测方法的灵敏性、准确性及可靠性。方法将微透析引导管置入正常成年大鼠纹状体内1周后,采用脑微透析技术动态收集2:00,6:00,10:00,14:00,18:00和22:00的透析液,并于次日22:00行尾静脉取血;用HPLC-荧光法和ELISA法分别测定透析液及血清样本中的MLT水平。结果 HPLC-荧光法测定MLT检出限为0.1pg/mL,在1.0~500pg/mL之间呈线性关系,回归方程y=10 446 x+8 603.5(r=0.998 7,P0.001),日间、日内精密度均6%;ELISA法检出限为5.92 pg/mL,检测线性范围为12.35~1 000pg/mL。用HPLC-荧光法测定2:00,6:00,10:00,14:00,18:00和22:00大鼠脑透析液中的MLT含量分别为(32.99±2.22),(18.25±2.27),(4.03±1.70),(2.92±1.08),(12.08±1.77)和(21.83±2.04)pg/mL,具有明显的昼夜节律性,探针回收率为12%~13%;22:00血清中的MLT含量为(82.62±8.09)pg/mL,提取回收率90%,且血清与透析液中的MLT水平呈高度正相关(r=0.987,P0.001);用ELISA法测定血清中的MLT含量为(81.31±9.62)pg/mL,且用2种方法测定血清中MLT差别无统计学意义(P=0.115)。结论用HPLC-荧光法测定大鼠脑微透析液和血清中的MLT含量具有灵敏、快速、准确、重复性好等特点,且HPLC-荧光法和ELISA法测定血清中的MLT具有较好的一致性。
[Abstract]:Objective to determine the content of melatonin (MLT) in adult rat brain microdialysis solution and serum by (HPLC) fluorescence and enzyme linked immunosorbent assay (ELISA), and to compare the sensitivity, accuracy and reliability of the two methods. Methods after placing the microdialysis tube into the striatum of the normal adult rats for 1 week, the hemodialysis solution of 2: 00 0: 6: 10: 00 10: 00: 14: 14: 18: 00 and 22:00 was collected dynamically by cerebral microdialysis technique, and the blood was taken from the tail vein at 22:00 the next day. The levels of MLT in dialysate and serum samples were measured by HPLC- fluorescence assay and ELISA method respectively. Results the detection limit of MLT by HPLC- fluorescence method was 0.1pg / mL, and the linear relationship between 1.0~500pg/mL was observed. The regression equation was 10 446 x 8 603.5 (r = 0.998 7). The detection limit of Elisa was 5.92 pg/mL, and the linear range was 12.351000 PG / mL. The contents of MLT in the dialysate of 2: 00 6: 10: 10: 00 and 22:00 were (32.99 卤2.22), (18.25 卤2.27), (4.03 卤1.70), (2.92 卤1.08), (12.08 卤1.77) and (21.83 卤2.04) pg/mL, respectively. The recovery rate of MLT in serum was (82.62 卤8.09) pg/mL, and 90%, and there was a high positive correlation between the MLT level in serum and dialysate (r = 0.987, P 0.001), the content of MLT in serum by ELISA method was (81.31 卤9.62) pg/mL, and there was no significant difference between the two methods (P = 0.115). Conclusion the determination of MLT in rat brain microdialysis solution and serum by HPLC- fluorescence method is sensitive, rapid, accurate and reproducible, and HPLC- fluorescence method and ELISA method have good consistency in the determination of MLT in serum.
【作者单位】： 福建医科大学神经生物学中心;中国疾病预防控制中心职业卫生与中毒控制所化学污染与健康安全重点实验室;
【基金】：国家自然科学基金(81241047) 福建省科技计划重点项目(2012Y0031)
【分类号】：R927.2;O657.7
[Abstract]:Objective to determine the content of melatonin (MLT) in adult rat brain microdialysis solution and serum by (HPLC) fluorescence and enzyme linked immunosorbent assay (ELISA), and to compare the sensitivity, accuracy and reliability of the two methods. Methods after placing the microdialysis tube into the striatum of the normal adult rats for 1 week, the hemodialysis solution of 2: 00 0: 6: 10: 00 10: 00: 14: 14: 18: 00 and 22:00 was collected dynamically by cerebral microdialysis technique, and the blood was taken from the tail vein at 22:00 the next day. The levels of MLT in dialysate and serum samples were measured by HPLC- fluorescence assay and ELISA method respectively. Results the detection limit of MLT by HPLC- fluorescence method was 0.1pg / mL, and the linear relationship between 1.0~500pg/mL was observed. The regression equation was 10 446 x 8 603.5 (r = 0.998 7). The detection limit of Elisa was 5.92 pg/mL, and the linear range was 12.351000 PG / mL. The contents of MLT in the dialysate of 2: 00 6: 10: 10: 00 and 22:00 were (32.99 卤2.22), (18.25 卤2.27), (4.03 卤1.70), (2.92 卤1.08), (12.08 卤1.77) and (21.83 卤2.04) pg/mL, respectively. The recovery rate of MLT in serum was (82.62 卤8.09) pg/mL, and 90%, and there was a high positive correlation between the MLT level in serum and dialysate (r = 0.987, P 0.001), the content of MLT in serum by ELISA method was (81.31 卤9.62) pg/mL, and there was no significant difference between the two methods (P = 0.115). Conclusion the determination of MLT in rat brain microdialysis solution and serum by HPLC- fluorescence method is sensitive, rapid, accurate and reproducible, and HPLC- fluorescence method and ELISA method have good consistency in the determination of MLT in serum.
【作者单位】： 福建医科大学神经生物学中心;中国疾病预防控制中心职业卫生与中毒控制所化学污染与健康安全重点实验室;
【基金】：国家自然科学基金(81241047) 福建省科技计划重点项目(2012Y0031)
【分类号】：R927.2;O657.7
【共引文献】
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1 李鑫宇;郭长虹;席绍峰;杨云;李焯原;刘家n，
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