A new phosphorimetry for the determination of trace alkaline phosphatase using multi-wall carbon nanotubes and its diagnosis of human diseases

Abstract

Multi-wall carbon nanotubes (MWNTs) could be modified as water soluble MWNTs (it was called as MWNTs-A and MWNTs-B) by chemical methods. MWNTs-A and MWNTs-B could emit room temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM). A new solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace alkaline phosphatase (ALP) was established based on the signal magnification effect of tween-80 and ALP on MWNTs-B's RTP intensity and the linear relationship between the content of ALP and the DeltaI(P) of the system. The linear range of this method was 0.0020-0.80 (fg spot(-1), sample volume: 0.40 microl spot(-1)), the regression equation of working curve was DeltaI(P) = 0.8170 + 96.84m(ALP) (fg spot(-1)), correlation coefficient (r) was 0.9986. This method had high sensitivity (detection limit (LD): 1.4 ag spot(-1)), good selectivity (Er < or = +/- 5 in-care-of, coexistence species were of no interference), high precision (RSDs were 4.4%-1.2%) and accuracy. It was applied to the determination of trace ALP in human serum and the diagnosis of human diseases. The results were tallied with those of enzyme-linked immunosorbent assay (ELISA). The mechanism of new SS-RTP for the determination of trace ALP was discussed, which laid the theory foundation for the analytical application of MWNTs in life science.