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A New Fluorescein Isothiocyanate‐Piperidine Self‐ordered Ring Phosphorescent Probe for the Determination of Trace Protein
Author(s) -
Lin Changqing,
Zhang Lihong,
Lin Shaoqin,
Su Limin,
Li Feiming,
Lin Liping,
Wang Xinxing,
Cai Jingping,
Liu Jiaming
Publication year - 2010
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.201090185
Subject(s) - chemistry , phosphorescence , detection limit , fluorescein isothiocyanate , bovine serum albumin , fluorescence , isothiocyanate , piperidine , analytical chemistry (journal) , linear range , chromatography , nuclear chemistry , stereochemistry , organic chemistry , physics , quantum mechanics
The COOH in fluorescein isothiocyanate (FITC) reacted with NH in piperidine (P) to form FITC‐P on the center of indentation of polyamide membrane (PAM) when drying for 2 min at (92±1)°C. Then, the FITC‐P diffused outward from the indentation center and formed the round SOR‐P‐FITC (containing the FITC‐P self‐ordered rings). Thus, multi‐FITC accumulated on SOR‐P‐FITC, leading to the enhancement of RTP signal on bio‐target, whose I p increased 2.0 times compared with non‐generated SOR. When bovine serum albumin (BSA) was added to the center of SOR‐P‐FITC, NCS of FITC in SOR‐P‐FITC reacted with NH 2 of BSA to form SOR‐P‐FITC‐BSA, which caused the RTP signal of FITC to enhance sharply. The Δ I p of the system was 3.4 times higher than that without β ‐CD and 4.0 times higher than that without SOR‐P‐FITC formed. Its Δ I p was linear to the content of BSA. Therefore, a new solid substrate‐room temperature phosphorimetry (SS‐RTP) for the determination of trace protein was established using SOR‐P‐FITC as a phosphorescent probe. Under the optimum condition, the linear range of this method was 0.040–16.0 ag·spot −1 with a detection limit (LD) of 8.5 zg·spot −1 (0.40 µL sample solution per spot, the corresponding concentration was 2.1×10 −17 g·mL −1 ), and the regression equation of working curve was Δ I p =3.848+4.240 m BSA (ag·spot ‐1 ), n =6, correlation coefficient ( r ) was 0.9993. This method with high sensitivity had been applied to determining the content of trace protein in the water samples, and the results coincided well with those obtained with pyrocatechol violet‐Mo(VI) method (P.V.M.M.). At the same time, the mechanism of SS‐RTP using SOR‐P‐FITC as a phosphorescent probe (SOR‐P‐FITC‐SS‐RTP) was discussed.