尿中高香草酸和香草扁桃酸测定的高效液相色谱-串联质谱法.pdfVIP

中华劳动卫生职业病杂志2016 年6 月第 34 卷第6 期 Chin J lnd Hyg Occup Dis , Jun 2016 ,Vo1. 34 ， Nι6 461 ·监测与检验技术· 尿中高香草酸和香草扁桃酸测定的高效液相 色谱-串联质谱法 李惠玲王国才未钧 [摘要] 目的 建立尿中高香草酸和香草扁桃酸测定的高效液相色谱-串联质谱(HPLC/M S/MS ) 法。方法取尿样2∞μl 置于离心管中，分别加入 20 f.Ll 氯瞠沙宗内标，混匀后加入 100μl 甲醇(含 1% 乙酸)，涡旋震荡 1 min ， 13 ∞o r/min(半径8.2 cm)离心 5 min，提取上清后，取 10μl 进样，利用 HPLC/MSI MS 法对高香草酸和香草扁桃酸进行定性定量分析。结果该方法测定高香草酸和香草扁挑酸的线性范 围分别为 20-10 ∞0 和 20-75∞ nglml ，相关系数分别为 0.9956 和 0.9970 ，检出限分别为 4.69 和 2 .33 nglml ，回收率分别为 85.72%-117.959品和 95.82%-113.78%。结论该方法操作简便、快速、准确、灵 敏度高，可满足尿中高香草酸和香草扁桃酸测定的要求。 [关键词] 色谱法，高压液相;尿;高香草酸;香草扁桃酸 Determination of HVA and VMA in urine using high peñormance Iiquid chromaωgraphy-tandem mωs spectrometη Li H uiling*, Wang Gωcω， Zhu }un. *Depanment olOccupationol Diseωe and Toxicatωn ， Chωyang Hospit a1 Affiliated ω the Capita1 Un加ersity 01Medic a1 Science , Beijing 1ω020 ， ChiTU1 Corresponding author: Zhu }un(E-mail:2584422151@) [Abstract] Objective To establish a method for the determination of homovanillic acid (HVA) and vanil mandelic acid (VMA) in urine , using high perfo口nance liquid chromatography-tandem mass spectromet可 (HPLC/M S/M S )detection. Meth叫s A 200 f.Ll volume of urine sample added with 20 f.Ll chlorzoxazone intemal standard was mixed in a centrifuge tube , then added 1 ∞μL methanol (containing 1 % acetic acid) , vortex mixed and centrifuged at 13 000 rpm for 1 min. A 10μ1 volume of the above supematant solution was injected into HPLC/M S/MS system for quantitative analysis. Results With intemal standard method , the calibration cu凹es of HVA and VMA were linear in the range of 20 -10 0∞ n!ifml and 20-7 500 n !ifml , with correlation coefficient of 0.995 6 and 0.997 0 , respectively. The LOQ of HVA and VMA was 4.69 nglml and 2 .33 nglml , respectively. The method recoveries were in the range of 85.729毛-117.959毛 for HVA , and 9