随机尿游离型甲氧基肾上腺素类物质的电化学检测新方法及临床应用

发布时间：2018-05-14 02:27

本文选题：丝网印刷碳电极 + 甲氧基肾上腺素类物质　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:1.建立一种简单、低廉测定随机尿游离型甲氧基肾上腺素类物质(metanephrines,MNs)的直接电化学检测方法。2.用所建立的方法测定嗜铬细胞瘤患者和对照组随机尿液游离型MNs浓度水平,同时也用高效液相色谱-电化学法(high performance liquid chromatography with electrochemical detection,HPLC-ED)检测所有研究对象血浆游离型MNs,并探讨MNs对嗜铬细胞瘤诊断效能。方法:1.分别采用循环伏安法(cyclic voltammetry,CV)和示差脉冲伏安法(differential pulse voltammetry,DPV)对MNs进行定性和定量分析。电极为丝网印刷碳电极(screen-printed carbon electrode,SPCE):工作电极和对电极由碳浆印制,参比电极为Ag/Ag Cl电极。2.标本采用固相萃取技术对其进行预处理后,加入PBS溶液中混匀,然后取50μL滴加于丝网印刷电极工作区上,并于连接电化学工作站用DPV进行反应信号测定。3.MNs水平与HPLP-ED法测定结果进行相关性分析。4.用建立方法测定对照组(n=63)和嗜铬细胞瘤病人组(n=22)随机尿中游离型MNs水平和HPLC-ED法测定血浆游离型MNs浓度,受试者工作曲线(receiver operating characteristic,ROC)用于评价对嗜铬细胞瘤诊断效能。结果:1.随机尿MNs的线性范围为0.25~12.5 mg/L,回归方程Y=0.916X-0.127,相关系数r=0.994,最低定量限是0.20 mg/L,最低检测限为0.05 mg/L。天内相对标准偏差和天间相对标准偏差分别在5.9%~9.7%和6.8%~9.9%之间。天内回收率为82.0%~101.1%,天间回收率为81.0%~96.5%。2.嗜铬细胞瘤患者组随机尿中游离型MNs水平分别采用建立方法与HPLC-ED法测定结果进行相关性分析,相关性良好(r=0.941,p=0.000)。3.本实验建立的方法检测随机尿游离型MNs和HPLC-ED法检测血浆中游离型MN和NMN浓度,嗜铬细胞瘤患者组较对照组显著增高(P均0.05),当血浆游离NMN和MN截断值分别为0.540 mg/L和0.128 mg/L,NMN的灵敏度与特异度分别是90.9%、95.2%,曲线下面积是0.975;MN的灵敏度与特异度分别是86.4%、85.7%,曲线下面积是0.916。本实验建立方法检测随机尿游离型MNs浓度截断值取0.404mg/L时,灵敏度与特异度分别是95.5%、92.4%,曲线下面积是0.981。结论:1.免修饰SPCE直接检测随机尿液中游离型MNs成本低廉、速度快。SPCE能进行批量生产,在临床诊断方面具有良好的应用前景。2.随机尿游离型MNs诊断PHEO灵敏度优于血浆游离型NMN和MN。基于随机尿具有取样方便、无创伤等优点,随机尿游离型MNs适用于嗜铬细胞瘤早期诊断和预后评估。
[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.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R736.6

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