超高效液相色谱-串联质谱法同时检测尿中三聚氰胺及其同系物  

Determination of four melamine and its derivatives in urine by ultra-performance liquid chromatography-tandem mass spectrometry

作　　者：刘少杰 黄菲菲 杨其帆 厉曙光 何更生[1] 陈波[1] 吴岷[1] Liu Shaojie;Huang Feifei;Yang Qifan;Li Shuguang;He Gengsheng;Chen Bo;Wu Min(Department of Nutrition and Food Hygiene,School of Public Health,Fudan University,Shanghai 200032,China;Chemical Laboratory,Jing*an District Center for Disease Control and Prevention,Shanghai 200041,China)

机构地区：[1]复旦大学公共卫生学院营养与食品卫生教研室,上海200032 [2]上海市静安区疾病预防控制中心理化检验科,上海200041

出　　处：《卫生研究》2022年第1期85-90,98,共7页Journal of Hygiene Research

摘　　要：目的采用超高效液相色谱-串联质谱法建立同时检测尿中三聚氰胺(melamine, MEL)及其同系物三聚氰酸二酰胺(ammeline, AMN)、三聚氰酸一酰胺(ammelide, AMD)和三聚氰酸(cyanuric acid, CYA)的方法。方法尿样加入内标后,经乙腈涡旋混匀,超声提取、离心,分别采用混合型阳离子交换或混合型阴离子交换固相萃取柱富集,经5%氨化甲醇或2%甲酸甲醇洗脱,40℃下氮气吹干,复溶混合后经Amide色谱柱(2.1 mm×100 mm, 1.7μm)分离,串联质谱仪在正负离子同时扫描模式下定性定量分析。结果三聚氰胺及其同系物的色谱峰均达到基线分离,峰型较好。MEL、AMN、AMD及CYA的检出限分别为0.03、0.04、0.04及0.05 ng/mL,定量限分别为0.11、0.12、0.14和0.15 ng/mL,基质加标样(浓度为0.2、5、50和200 ng/mL)的回收率为94.6%~106.5%,日内精密度为0.73%~8.34%,日间精密度均小于6.80%(n=6)。AMD和CYA在低浓度时主要表现为基质增强效应,而AMN随着浓度的增加呈现出基质抑制效应。对60份随机成人尿样进行检测,发现MEL及其同系物的总检出率为75.0%,且4种物质的平均浓度男性均高于女性。结论利用超高效液相色谱-串联质谱法可同时测定尿中MEL及其同系物AMN、AMD和CYA,方法检出限和定量限较低,准确性和精密度较好。OBJECTIVE To establish a method for simultaneous detection of melamine(MEL), ammeline(AMN), ammelide(AMD) and cyanuric acid(CYA) in urine by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). METHODS After the urine sample was added to the internal standard, they were mixed by acetonitrile, ultrasonic extraction and centrifugation, enriched by MCX or MAX solid phase extraction column, eluted with 5% ammoniated methanol or 2% formic acid methanol, dried under nitrogen at 40 ℃, separated by Amide chromatography column(2.1 mm×100 mm, 1.7 μm), qualitatively and quantitatively analyzed by mass spectrometer in the simultaneous scanning mode of positive and negative ions. RESULTS Melamine and its derivatives were completely separated with great chromatography peak. The limit of detection(LOD) for melamine, ammeline, ammelide and cyanuric acid were 0.03, 0.04, 0.04 and 0.05 ng/mL, and the limit of quantification(LOQ) were 0.11, 0.12, 0.14 and 0.15 ng/mL, respectively. The recovery rate of standard addition samples of 0.2, 5, 50 and 200 ng/mL ranged from 94.6% to 106.5%, the intra-day precision was between 0.73% and 8.34%, and the inter-day precision was less than 6.80%(n=6). At low concentrations, AMD and CYA mainly showed matrix enhancing effect, while AMN showed matrix inhibitory effect with increasing concentration. The overall detection rate of melamine and its derivatives was 75.0% in 60 parcels of random urine. The average concentration of urinary melamine and its derivatives were higher in men. CONCLUSION UPLC-MS/MS can simultaneously determine four melamine and its derivatives in urine. The LOD and LOQ of the current method were low with great accuracy and precision.

关 键 词：三聚氰胺 三聚氰酸 尿液 生物监测 超高效液相色谱-串联质谱法 

分 类 号：R155.5[医药卫生—营养与食品卫生学]