随机尿游离型NMN、MN HPLC-ED检测方法的建立和临床应用  

Method establishment of spot urine free NMN and MN by HPLC-ED and its clinical application

作　　者：张磊[1] 左明 邹文毕 王浩然 张晓清[3] 丁敏[3] ZHANG Lei;ZUO Ming;ZOU Wenbi;WANG Haoran;ZHANG Xiaoqing;DING Min(Department of Clinical Laboratory,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Nuclear Medicine Laboratory,the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Key Laboratory of Clinical Laboratory Diagnostics,Ministry of Education,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学附属第一医院医学检验科,重庆400016 [2]重庆医科大学附属第二医院核医学科实验室,重庆400016 [3]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016

出　　处：《检验医学》2021年第5期524-529,共6页Laboratory Medicine

摘　　要：目的建立检测随机尿游离型甲氧基去甲肾上腺素(NMN)、甲氧基肾上腺素(MN)的高效液相色谱-电化学(HPLC-ED)方法,并探讨其临床应用价值。方法完善样本处理流程及色谱条件,建立检测随机尿游离型NMN、MN的HPLC-ED法。采用HPLC-ED法检测35例嗜铬细胞瘤患者(嗜铬细胞瘤组)、70例原发性高血压患者(高血压组)、78例非嗜铬细胞瘤肾上腺占位患者(肾上腺占位组)和44名体检健康者(正常对照组)的随机尿和血浆游离型NMN、MN浓度,同时检测肌酐(Cr),计算尿NMN/Cr比值和尿MN/Cr比值。采用受试者工作特征(ROC)曲线评估各项指标诊断嗜铬细胞瘤的效能。结果HPLC-ED法检测随机尿游离型NMN、MN的线性范围均为25~2500μg/L,检测限[信噪比(RSN)=3]分别为5.9和11.3μg/L,平均回收率为92.8%~103.4%。NMN高、低浓度的批内变异系数(CV)分别为3.7%、2.2%,批间CV分别为5.6%、3.3%;MN高、低浓度批内CV分别为4.3%、2.7%,批间CV分别为5.1%、3.4%。嗜铬细胞瘤组随机尿游离型NMN、MN浓度和尿NMN/Cr比值、尿MN/Cr比值明显高于高血压组、肾上腺占位组及正常对照组(P<0.001),高血压组、肾上腺占位组及正常对照组之间差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,随机尿游离型NMN、随机尿游离型MN、尿NMN/Cr比值、尿MN/Cr比值、血浆游离型NMN、血浆游离型MN诊断嗜铬细胞瘤的曲线下面积(AUC)分别为0.91、0.88、0.96、0.94、0.93和0.88。嗜铬细胞瘤患者术后1周尿MN/Cr比值均降为正常,但有2例患者术后血浆MN浓度高于术前,有1例患者血浆NMN浓度高于术前。结论建立了检测随机尿游离型NMN、MN浓度的HPLC-ED方法。尿NMN/Cr比值和尿MN/Cr比值诊断嗜铬细胞瘤的效能均优于血浆游离型NMN、MN,且在嗜铬细胞瘤的术后评估中也有较好的价值。Objective To establish a method for spot urine free normetanephrine(NMN)and metanephrine(MN)by high performance liquid chromatography-electrochemical detection(HPLC-ED),and to investigate the clinical application role.Methods Sample processing and chromatographic conditions were improved,and HPLC-ED for spot urine free NMN and MN was established.HPLC-ED was used to determine NMN and MN in 35 patients with pheochromocytoma,70 patients with primary hypertension,78 patients with adrenal mass but without pheochromocytoma and 44 healthy subjects.Creatinine(Cr)was determined as well.NMN/Cr ratio and MN/Cr ratio were calculated.Receiver operating characteristic(ROC)curve was used to evaluate the efficiency for diagnosing pheochromocytoma.Results The linear range of HPLC-ED for the determinations of NMN and MN was 25-2500μg/L.The determination limits[signal-to-noise ratio(RSN)=3]were 5.9 and 11.3μg/L,respectively.The average recovery was 92.8%-103.4%.The within-run coefficients of variation(CV)of high-concentration and low-concentration NMN were 3.7%and 2.2%,and the between-run CV of high-concentration and low-concentration NMN were 5.6%and 3.3%,respectively.The within-run CV of high-concentration and low-concentration MN were 4.3%and 2.7%,and the between-run CV of high-concentration and low-concentration MN were 5.1%and 3.4%,respectively.The concentrations of NMN and MN,NMN/Cr ratio and MN/Cr ratio in pheochromocytoma group were higher than those in hypertension group,adrenal space-occupying group and healthy control group(P<0.001),and there was no statistical significance among hypertension group,adrenal space-occupying group and healthy control group(P>0.05).ROC curve analysis showed that the area under curve(AUC)in diagnosing pheochromocytoma of urine free NMN,urine free MN,NMN/Cr ratio,MN/Cr ratio,plasma free NMN and plasma free NMN were 0.91,0.88,0.96,0.94,0.93 and 0.88,respectively.MN/Cr ratio in urine decreased to normal in 1 week after operation in pheochromocytoma patients,but the concentration of plasma MN i

关 键 词：甲氧基去甲肾上腺素 甲氧基肾上腺素 高效液相色谱-电化学法 嗜铬细胞瘤 

分 类 号：R446.1[医药卫生—诊断学]