本文選題：絲網(wǎng)印刷碳電極 + 甲氧基腎上腺素類物質(zhì)��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:1.建立一種簡(jiǎn)單、低廉測(cè)定隨機(jī)尿游離型甲氧基腎上腺素類物質(zhì)(metanephrines,MNs)的直接電化學(xué)檢測(cè)方法。2.用所建立的方法測(cè)定嗜鉻細(xì)胞瘤患者和對(duì)照組隨機(jī)尿液游離型MNs濃度水平,同時(shí)也用高效液相色譜-電化學(xué)法(high performance liquid chromatography with electrochemical detection,HPLC-ED)檢測(cè)所有研究對(duì)象血漿游離型MNs,并探討MNs對(duì)嗜鉻細(xì)胞瘤診斷效能。方法:1.分別采用循環(huán)伏安法(cyclic voltammetry,CV)和示差脈沖伏安法(differential pulse voltammetry,DPV)對(duì)MNs進(jìn)行定性和定量分析。電極為絲網(wǎng)印刷碳電極(screen-printed carbon electrode,SPCE):工作電極和對(duì)電極由碳漿印制,參比電極為Ag/Ag Cl電極。2.標(biāo)本采用固相萃取技術(shù)對(duì)其進(jìn)行預(yù)處理后,加入PBS溶液中混勻,然后取50μL滴加于絲網(wǎng)印刷電極工作區(qū)上,并于連接電化學(xué)工作站用DPV進(jìn)行反應(yīng)信號(hào)測(cè)定。3.MNs水平與HPLP-ED法測(cè)定結(jié)果進(jìn)行相關(guān)性分析。4.用建立方法測(cè)定對(duì)照組(n=63)和嗜鉻細(xì)胞瘤病人組(n=22)隨機(jī)尿中游離型MNs水平和HPLC-ED法測(cè)定血漿游離型MNs濃度,受試者工作曲線(receiver operating characteristic,ROC)用于評(píng)價(jià)對(duì)嗜鉻細(xì)胞瘤診斷效能。結(jié)果:1.隨機(jī)尿MNs的線性范圍為0.25~12.5 mg/L,回歸方程Y=0.916X-0.127,相關(guān)系數(shù)r=0.994,最低定量限是0.20 mg/L,最低檢測(cè)限為0.05 mg/L。天內(nèi)相對(duì)標(biāo)準(zhǔn)偏差和天間相對(duì)標(biāo)準(zhǔn)偏差分別在5.9%~9.7%和6.8%~9.9%之間。天內(nèi)回收率為82.0%~101.1%,天間回收率為81.0%~96.5%。2.嗜鉻細(xì)胞瘤患者組隨機(jī)尿中游離型MNs水平分別采用建立方法與HPLC-ED法測(cè)定結(jié)果進(jìn)行相關(guān)性分析,相關(guān)性良好(r=0.941,p=0.000)。3.本實(shí)驗(yàn)建立的方法檢測(cè)隨機(jī)尿游離型MNs和HPLC-ED法檢測(cè)血漿中游離型MN和NMN濃度,嗜鉻細(xì)胞瘤患者組較對(duì)照組顯著增高(P均0.05),當(dāng)血漿游離NMN和MN截?cái)嘀捣謩e為0.540 mg/L和0.128 mg/L,NMN的靈敏度與特異度分別是90.9%、95.2%,曲線下面積是0.975;MN的靈敏度與特異度分別是86.4%、85.7%,曲線下面積是0.916。本實(shí)驗(yàn)建立方法檢測(cè)隨機(jī)尿游離型MNs濃度截?cái)嘀等?.404mg/L時(shí),靈敏度與特異度分別是95.5%、92.4%,曲線下面積是0.981。結(jié)論:1.免修飾SPCE直接檢測(cè)隨機(jī)尿液中游離型MNs成本低廉、速度快。SPCE能進(jìn)行批量生產(chǎn),在臨床診斷方面具有良好的應(yīng)用前景。2.隨機(jī)尿游離型MNs診斷PHEO靈敏度優(yōu)于血漿游離型NMN和MN�；陔S機(jī)尿具有取樣方便、無創(chuàng)傷等優(yōu)點(diǎn),隨機(jī)尿游離型MNs適用于嗜鉻細(xì)胞瘤早期診斷和預(yù)后評(píng)估。
[Abstract]:Purpose 1. A simple and low cost direct electrochemical method for the determination of free methoxyadrenaline (MNs) in random urine was established. The level of free MNs in urine of patients with pheochromocytoma and control group was determined by the established method. Plasma free MNs were also detected by high performance liquid chromatography-electrochemical method (HPLC-EDD) in all subjects, and the diagnostic efficacy of MNs for pheochromocytoma was investigated. Method 1: 1. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were used to analyze MNs qualitatively and quantitatively. The working and opposite electrodes are printed by carbon paste, and the reference electrode is Ag/Ag Cl electrode. The sample was pretreated by solid phase extraction (SPE) and mixed in PBS solution. Then 50 渭 L was added to the screen printing electrode working area. The correlation between the level of MNs and the results of HPLP-ED method was analyzed with DPV. 3. The levels of free type MNs in random urine and plasma free type MNs were measured by HPLC-ED method. The receiver operating character roc was used to evaluate the diagnostic efficacy of pheochromocytoma in patients with pheochromocytoma. The result is 1: 1. The linear range of random urinary MNs was 0.25 渭 g / L, the regression equation was 0.916X-0.127, the correlation coefficient was 0.994, the lowest quantitative limit was 0.20 mg / L, and the lowest detection limit was 0.05mg / L. The relative standard deviations within days and between days were 5.9% and 6.9%, respectively. The recovery rate was 82.0% 101.1% in days and 81.0% 96.5% in days. The level of free MNs in random urine of patients with pheochromocytoma was analyzed by establishing method and HPLC-ED method. The correlation was good. The established method was used to detect the concentration of free MN and NMN in plasma by random urinary free MNs and HPLC-ED. The sensitivity and specificity of plasma free NMN and MN were 0.540 mg/L and 0.128 mg / L NMN, respectively. The sensitivity and specificity of plasma free NMN and MN were 90.975 and 0.975 MN, respectively. The area under the curve was 0.975 MN, and the area under the curve was 0.916. In this study, the sensitivity and specificity of random urinary free MNs were 95.5and 92.42.The area under the curve was 0.981.When the truncation value of random urinary free MNs was measured by 0.404mg/L, the sensitivity and specificity of the method were 95.5and 92.4th.The area under the curve was 0.981. Conclusion 1. The direct detection of free MNs in random urine by non-modified SPCE has low cost and high speed. It can be used in batch production and has a good application prospect in clinical diagnosis. The sensitivity of random urinary free MNs in the diagnosis of PHEO was superior to that of plasma free NMN and MNs. Random urine free MNs is suitable for early diagnosis and prognosis evaluation of pheochromocytoma.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R736.6

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