Coupling technique of self-ordered ring and phosphorimetry for the determination of alkaline phosphatase and diseases prediction

Rhodamine S could emit strong and stable room temperature phosphorescence (RTP) on polyamide membrane (PAM) in the presence of heavy atom perturber Pb2+. When Rhodamine S-piperidine solution was dropped on PAM, the red (Rhod.S)n-P-SOR (Rhod.S, (Rhod.S)n, P and SOR refer to alizarin red S, multiple Rhod.S molecules, piperidine and self-ordered ring, respectively) formed on PAM, leading to the enhancement of room temperature phosphorimetry (RTP) intensity (Ip, 117.2) of (Rhod.S)n-P-SOR system, which was 2.4 times higher than that without SOR (Ip, 48.1). Wheat germ agglutinin (WGA) was labelled with (Rhod.S)n-P-SOR by the –NH– of Rhod.S reacting with the –COOH of WGA to form WGA-(Rhod.S)n-P-SOR. The formation of WGA-AP-WGA-(Rhod.S)n-P-SOR in the affinity adsorption (AA) reaction carried out between the –COOH of WGA in WGA-(Rhod.S)n-P-SOR and the –NH2 of alkaline phosphatase (AP) caused the RTP intensity (ΔIp) of the WGA-AP-WGA-(Rhod.S)n-P-SOR system 7.8 times larger than that without (Rhod.S)n-P-SOR. Therefore, the coupling technique of SOR and solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace AP has been established. This method possessed good selectivity, high sensitivity (Detection limit (L.D) was 3.4 × 10−16 g mL−1) and accuracy, and it has been applied to the determination of trace AP in human serum and the forecast of human diseases, and the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Besides, the mechanism of the coupling technique for the determination of AP was discussed.

Graphical abstractCoupling technique of self-ordered ring and phosphorimetry for the determination of trace alkaline phosphatase (AP) and forecast human diseases has been established based on affinity adsorption reaction between Rhodamine S-piperidine-self-ordered ring-wheat germ agglutinin ((Rhod.S)n-P-SOR-WGA) and AP. This method has been applied to the determination of trace AP in human serum and the forecast of human diseases, and the results agreed well with those obtained by enzyme-linked immunoassay. Besides, the mechanism of the coupling technique for the determination of AP was discussed.Figure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Phosphorescence reagent (Rhod.S)n-P-SOR was developed by self-ordered ring. ► Phosphorimetry for determination of alkaline phosphatase (AP) was established. ► The method could be applied to the detection of AP and the forecast of diseases. ► Mechanism of SOR and phosphorimetry for the determination of AP was discussed. ► Immunoassay, coupling technique of SOR and phosphorimetry, lectin were developed.