A New Fluorescein Isothiocyanate-Piperidine Self-ordered Ring Phosphorescent Probe for the Determination of Trace Protein

作者：Lin Changqing; Zhang Lihong; Lin Shaoqin; Su Limin; Li Feiming; Lin Liping; Wang Xinxing; Cai Jingping; Liu Jiaming^*

来源：Chinese Journal of Chemistry, 2010, 28(6): 997-1004.

• fluorescein isothiocyanate-piperidine self-ordered ring
• phosphorescent probe
• solid substrate-room temperature phosphorimetry
• protein
• inclusion compound

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) degrees 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 Delta I(p) of the system was 3.4 times higher than that without beta-CD and 4.0 times higher than that without SOR-P-FITC formed. Its Delta 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 center dot spot(-1) with a detection limit (LD) of 8.5 zg center dot spot(-1) (0.40 mu L sample solution per spot, the corresponding concentration was 2.1 x 10(-17) g center dot mL(-1)), and the regression equation of working curve was Delta I(p)=3.848+4.240m(BSA) (ag center dot 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.

• 出版日期2010-6
• 单位福建教育学院; 闽南师范大学