Development of a nano-SiO2based enzyme-linked ligand binding assay for the determination of ibuprofen in human urine.
The application domains of classic enzyme-linked ligand binding assay (ELBA) is relatively narrow due to the high cost and hardly available binding receptor. In here, we described for the first time the possibility of developing a new ELBA based on silica nanoparticles (nano-SiO2) to assess the ibuprofen in human urine. Nano-SiO2with a large surface area was introduced as stationary phase to improve the analytical performance. In the experiment, a competitively binding procedure with human serum albumin (HSA) was performed between the ibuprofen presented in sample and horseradish peroxidase labeled ibuprofen (HRP-ibuprofen) subsequently added. After centrifugal separation, the HRP/ibuprofen/nano-SiO2composite catalyzed the substrate solution (TMB/H2O2) with a color change from colorless to yellow for quantitative measurement via an ultraviolet spectrophotometer. As a validation of the new principle, the developed nano-ELBA method was applied in the determination of ibuprofen excreted in human urine with excellent performance. This detection range only depends on the solubility of ligand and sensitivity of UV spectrophotometer. Our results indicate that this new method demonstrated to be able to rapidly and adequately determine the concentration of components in biological samples and advocate its effectiveness for various applications.
CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization & Ecological Restoration and Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
Qian-Long
QL
Wang
State Key Laboratory for Quality Research in Chinese Medicines, Macau University of Science and Technology, Macau, China; School of Pharmacy, Chengdu Medical College, Chengdu 610083, China.
Jing
J
Xie
CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization & Ecological Restoration and Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China; Key Laboratory of Natural Medicine and Translational Medicine, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
Xing-De
XD
Li
CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization & Ecological Restoration and Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China; Key Laboratory of Natural Medicine and Translational Medicine, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
Li-Sheng
LS
Ding
CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization & Ecological Restoration and Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China; Key Laboratory of Natural Medicine and Translational Medicine, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
Jian
J
Liang
State Key Laboratory for Quality Research in Chinese Medicines, Macau University of Science and Technology, Macau, China. Electronic address: pluo@must.edu.mo.
Pei
P
Luo
Key Laboratory of Natural Medicine and Translational Medicine, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China. Electronic address: qingls@cib.ac.cn.
Lin-Sen
LS
Qing
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000652:urine*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007052:Ibuprofen / Q000378:metabolism / Q000652:urine*; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism; D000075462:Serum Albumin, Human / Q000737:chemistry / Q000378:metabolism*; D012822:Silicon Dioxide / Q000737:chemistry* / Q000378:metabolism
D000894:Anti-Inflammatory Agents, Non-Steroidal; D012822:Silicon Dioxide; D006735:Horseradish Peroxidase; D007052:Ibuprofen; D000075462:Serum Albumin, Human
Direct competitive; Enzyme-linked ligand binding assay; HSA; Ibuprofen; Nano-SiO(2)