【摘要】：目的采用高效液相色譜(HPLC)-熒光法和酶聯(lián)免疫(ELISA)法測(cè)定成年大鼠腦微透析液及血清中的褪黑素(MLT)含量,比較2種檢測(cè)方法的靈敏性、準(zhǔn)確性及可靠性。方法將微透析引導(dǎo)管置入正常成年大鼠紋狀體內(nèi)1周后,采用腦微透析技術(shù)動(dòng)態(tài)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行尾靜脈取血;用HPLC-熒光法和ELISA法分別測(cè)定透析液及血清樣本中的MLT水平。結(jié)果 HPLC-熒光法測(cè)定MLT檢出限為0.1pg/mL,在1.0~500pg/mL之間呈線性關(guān)系,回歸方程y=10 446 x+8 603.5(r=0.998 7,P0.001),日間、日內(nèi)精密度均6%;ELISA法檢出限為5.92 pg/mL,檢測(cè)線性范圍為12.35~1 000pg/mL。用HPLC-熒光法測(cè)定2:00,6:00,10:00,14:00,18:00和22:00大鼠腦透析液中的MLT含量分別為(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,具有明顯的晝夜節(jié)律性,探針回收率為12%~13%;22:00血清中的MLT含量為(82.62±8.09)pg/mL,提取回收率90%,且血清與透析液中的MLT水平呈高度正相關(guān)(r=0.987,P0.001);用ELISA法測(cè)定血清中的MLT含量為(81.31±9.62)pg/mL,且用2種方法測(cè)定血清中MLT差別無(wú)統(tǒng)計(jì)學(xué)意義(P=0.115)。結(jié)論用HPLC-熒光法測(cè)定大鼠腦微透析液和血清中的MLT含量具有靈敏、快速、準(zhǔn)確、重復(fù)性好等特點(diǎn),且HPLC-熒光法和ELISA法測(cè)定血清中的MLT具有較好的一致性。
[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 鹵2.22), (18.25 鹵2.27), (4.03 鹵1.70), (2.92 鹵1.08), (12.08 鹵1.77) and (21.83 鹵2.04) pg/mL, respectively. The recovery rate of MLT in serum was (82.62 鹵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 鹵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.
【作者單位】： 福建醫(yī)科大學(xué)神經(jīng)生物學(xué)中心;中國(guó)疾病預(yù)防控制中心職業(yè)衛(wèi)生與中毒控制所化學(xué)污染與健康安全重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(81241047) 福建省科技計(jì)劃重點(diǎn)項(xiàng)目(2012Y0031)
【分類號(hào)】：R927.2;O657.7

【共引文獻(xiàn)】

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1 李鑫宇;郭長(zhǎng)虹;席紹峰;楊云;李焯原;劉家n，

本文編號(hào)：2279262