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Determination of Trace Alkaline Phosphatase by Solid‐Substrate Room‐Temperature Phosphorimetry Based on Triticum vulgare Lectin Labeled with Fullerenol
Author(s) -
Liu JiaMing,
Gao Fei,
Huang HongHua,
Zeng LiQing,
Huang XiaoMei,
Zhu GuoHui,
Li ZhiMing
Publication year - 2008
Publication title -
chemistry and biodiversity
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 70
eISSN - 1612-1880
pISSN - 1612-1872
DOI - 10.1002/cbdv.200890057
Subject(s) - chemistry , lectin , adsorption , substrate (aquarium) , detection limit , alkaline phosphatase , chromatography , analytical chemistry (journal) , nuclear chemistry , enzyme , biochemistry , organic chemistry , oceanography , geology
Fullerenol (F) shows a strong and stable room‐temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM) at λ $\rm{ _{{\rm{ex}}}^{\max } }$ / λ $\rm{ _{{\rm{em}}}^{\max } }$ =542.0/709.4 nm. When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal. It was also found that quantitative specific affinity‐adsorption reaction can be carried out between Triticum vulgare lectin (WGA) labeled with DBS‐F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA‐ALP‐WGA‐F‐DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the Δ I p value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity‐adsorption solid‐substrate room‐temperature phosphorimetry (AA‐SS‐RTP) was established, using fullerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot −1 (corresponding concentration: 2.8×10 −14 g ml −1 , namely 2.8×10 −16 mol l −1 ). This method with high sensitivity, accuracy, and precision has been successfully applied to the determination of the content of ALP in human serum survey and forecast human disease, and the results are tallied with those using alkaline phosphatase kits. The mechanism for the determination of ALP using AA‐SS‐RTP was also discussed.