药学学报, 2014, 49(4): 517-523
引用本文:
谢丽伟, 赵祥升, 孔维军, 王宇婷, 胡一晨, 欧阳臻, 杨美华. 分子印迹聚合物固相萃取柱净化-HPLC-FLD测定人体尿液中的赭曲霉毒素A[J]. 药学学报, 2014, 49(4): 517-523.
XIE Li-wei, ZHAO Xiang-sheng, KONG Wei-jun, WANG Yu-ting, HU Yi-chen, OU-YANG Zhen, YANG Mei-hua. Determination of ochratoxin A in human urine by HPLC-FLD after cleaned-up by molecularly imprinted polymer solid phase extraction column[J]. Acta Pharmaceutica Sinica, 2014, 49(4): 517-523.

分子印迹聚合物固相萃取柱净化-HPLC-FLD测定人体尿液中的赭曲霉毒素A
谢丽伟1,2, 赵祥升1,3, 孔维军1, 王宇婷1,2, 胡一晨1, 欧阳臻2, 杨美华1,3
1. 中国医学科学院、北京协和医学院药用植物研究所, 北京 100193;
2. 江苏大学药学院, 江苏 镇江 212013;
3. 中国医学科学院、北京协和医学院药用植物研究所海南分所, 海南 万宁 571533
摘要:
建立一种测定人体尿液赭曲霉毒素A(OTA)含量的分子印迹聚合物固相萃取柱净化、高效液相色谱-荧光检测(HPLC-FLD)方法。尿液样品在北京地区随机收集,用0.1 mol·L-1 HCl溶液调至pH 2.5,经分子印迹聚合物固相萃取柱净化后,HPLC-FLD检测其所含赭曲霉毒素A的含量,液相色谱-质谱法(LC-MS/MS)确证阳性样品。赭曲霉毒素A在0.75~250 ng·mL-1内线性关系良好。OTA在3个加样水平的回收率为90.6%~101.9%,RSD为0.1%~1.6%。65份志愿者尿液共检测出5份阳性样品,OTA最大含量为0.091 ng·mL-1。本文首次将分子印迹聚合物固相萃取柱应用于净化和富集人体尿液中的OTA,方法简单、快速、可靠,适用于人体尿液中OTA的检测。
关键词:    分子印迹聚合物固相萃取      高效液相色谱-荧光检测法      赭曲霉毒素A     
Determination of ochratoxin A in human urine by HPLC-FLD after cleaned-up by molecularly imprinted polymer solid phase extraction column
XIE Li-wei1,2, ZHAO Xiang-sheng1,3, KONG Wei-jun1, WANG Yu-ting1,2, HU Yi-chen1, OU-YANG Zhen2, YANG Mei-hua1,3
1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China;
2. School of Pharmacy, Jiangsu University, Zhenjiang 212013, China;
3. Hainan Branch Institute of Medicinal Plant Development, Chinese Academy of Medicinal Sciences and Peking Union Medical College, Wanning 571533, China
Abstract:
A method was developed for the determination of ochratoxin A (OTA) in human urine by HPLC-FLD after molecularly imprinted polymer solid phase extraction (MIP-SPE) column. After the pH being adjusted to 2.5 with 0.1 mol·L-1 HCl, sample was cleaned up with MIP-SPE column for ochratoxin A, the analyte was analyzed by high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD), and finally all the positive results were confirmed by LC-MS/MS. Recoveries from urine samples spiked with OTA at levels ranging from 2 to 20 ng·mL-1 were 90.6%-101.9%, and RSDs were 0.1%-1.6%. Sixty-five volunteers living in Beijing took part in the study, of which 5 were found containing OTA in their urine and the highest value was 0.091 ng·mL-1. The MIP-SPE column was firstly applied to purify and concentrate OTA in human urine, this method is simple, rapid and reliable and can be used to determine the contents of OTA in human urine.
Key words:    molecularly imprinted polymer solid phase extration    HPLC-FLD    ochratoxin A   
收稿日期: 2013-10-19
基金项目: 国家自然科学基金面上项目(81274072);协和青年科研基金和中央高校基本科研业务费专项资金资助项目(3332013078).
通讯作者: 杨美华
Email: yangmeihua15@hotmail.com
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