CAJ | 학술논문

目的：建立尿液儿茶酚胺[包括肾上腺素（E）、去甲肾上腺素（NE）和多巴胺（DA）]的液相色谱－串联质谱（LC－MS／MS）检测方法。方法以 E－d3、NE－d6和 DA－d4盐酸盐为内标，采用 WATERS ACQUITY UPLC? HSS T3色谱柱（2．1 mm ×100 mm，1．8μm）进行分离。流动相 A1为2‰甲酸水溶液，流动相 B1为甲醇（纯度为99．9％），流速为0．5 mL／min，按 A1∶B1＝95∶5等度洗脱。采用电喷雾（ESI）正离子模式进行质谱分析。对方法的线性、准确度（回收率）、不精密度及最低检出限等进行性能验证。选取表面健康人群65名，建立 E、NE 和 DA 的参考区间。结果采用建立的 LC－MS／MS 检测尿液 E、NE 和 DA，保留时间分别为0．58、0．89和0．63 min，检测范围为0．50～800．00 ng／mL。 E、NE 和 DA 的加标平均回收率分别为97．95％、97．78％和101．03％，最低检出限分别为0．25、2．50和2．50 ng／mL。 E、NE 和 DA 的参考区间分别为5．4～25．8、28．5～73．1和269．1～420．5μg／24 h。结论建立的LC－MS／MS 方法能同时检测尿液中的 E、NE 和 DA，可为嗜铬细胞瘤的诊断提供可靠的信息。
목적：건립뇨액인다분알[포괄신상선소（E）、거갑신상선소（NE）화다파알（DA）]적액상색보－천련질보（LC－MS／MS）검측방법。방법이 E－d3、NE－d6화 DA－d4염산염위내표，채용 WATERS ACQUITY UPLC? HSS T3색보주（2．1 mm ×100 mm，1．8μm）진행분리。류동상 A1위2‰갑산수용액，류동상 B1위갑순（순도위99．9％），류속위0．5 mL／min，안 A1∶B1＝95∶5등도세탈。채용전분무（ESI）정리자모식진행질보분석。대방법적선성、준학도（회수솔）、불정밀도급최저검출한등진행성능험증。선취표면건강인군65명，건립 E、NE 화 DA 적삼고구간。결과채용건립적 LC－MS／MS 검측뇨액 E、NE 화 DA，보류시간분별위0．58、0．89화0．63 min，검측범위위0．50～800．00 ng／mL。 E、NE 화 DA 적가표평균회수솔분별위97．95％、97．78％화101．03％，최저검출한분별위0．25、2．50화2．50 ng／mL。 E、NE 화 DA 적삼고구간분별위5．4～25．8、28．5～73．1화269．1～420．5μg／24 h。결론건립적LC－MS／MS 방법능동시검측뇨액중적 E、NE 화 DA，가위기락세포류적진단제공가고적신식。
Objective To establish a liquid chromatography-tandem mass spectrometry ( LC-MS/MS) for the determination of urinary catecholamines, including epinephrine ( E), norepinephrine ( NE) and dopamine ( DA). Methods E, NE and DA were separated by WATERS ACQUITY UPLC? HSS T3 (2.1 mm ×100 mm,1.8 μm column) using E-d3, NE-d6 and DA-d4 as internal standards.The mobile phase consisted of 2‰ formic acid aqueous solution ( A1) and methyl alcohol ( B1, purity =99.9%).The flow rate of 0.50 mL/min was applied to the chromatographic column.Flow starting was performed with A1 ∶B1 =95 ∶5.Electrospray ionization ( ESI) was operated in positive ion mode.The LC-MS/MS was used, with linearity, accuracy (recovery rate), imprecision and lower limit of determination being evaluated.A total of 65 healthy subjects were enrolled, and the reference ranges for E, NE and DA were established.Results The retention times of E, NE and DA were 0.58, 0.89, 0.63 min, respectively.The determination range was from 0.50 ng/mL to 800.00 ng/mL.The mean recovery ranges were 97.95%, deter mination range 97.78% and 101.03%.The lower limits of determination for E, NE and DA were 0.25, 2.50 and 2.50 ng/mL.The reference ranges were 5.4-25.8 μg/24 h for E, 28.5-73.1 μg/24 h for NE and 269.1-420.5 μg/24 h for DA.Conclusions The LC-MS/MS for the simultaneous determination of E, NE and DA has been established and is suitable for clinical application.It can provide reliable information for the diagnosis of phaeochromocytoma.