Ningning Zhu

Determination of hydroxyl radical by capillary zone electrophoresis with amperometric detection.

Hydroxyl radical (OH*) can cause severe damage to cells and tissues. However, its analysis is very difficult for its high reactivity and very short half-life. In this paper, a simple and highly sensitive method, capillary zone electrophoresis with amperometric detection (CZE-AD) was introduced indirectly to determine OH* by determining its reaction products with salicylic acid (SAL), 2,3-dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA). The optimum conditions of CZE-AD for the determination of 2,3-DHBA and 2,5-DHBA were explored. Under the optimum conditions, SAL, 2,3-DHBA and 2,5-DHBA could be perfectly separated within 15 min, and the linearity ranges of 2,3-DHBA and 2,5-DHBA were between 1.0 x 10(-7) and 1.0 x 10(-4) mol l(-1). Their detection limits were as low as 2 x 10(-8) mol l(-1), which were much lower than that in CE-UV method. The method was also applied to study the free OH* scavenging activity of angelica polysaccharide. The experimental results showed that this CZE-AD method was very sensitive and practical in both the determination of free OH* and the evaluation of free OH* scavenging activities of antioxidants.

PAMAM dendrimer-enhanced DNA biosensors based on electrochemical impedance spectroscopy

A novel, simple and sensitive DNA biosensor based on DNA-poly(amidoamine) (PAMAM) dendrimer nanoconjugates was developed by using the electrochemical impedance spectroscopy (EIS) technique. In this context, the assay relies on the hybridization of the single-stranded DNA (ssDNA) probe covalently conjugated on a mercaptoacetic acid self-assembled monolayer on gold electrodes, with the generation 4.5 (G-4.5) PAMAM-target DNA complex in solution. Once the double-stranded DNA (dsDNA) formed on the gold electrodes, G-4.5 PAMAM bearing carboxyls on the periphery was anchored on the hybrids; the changes of interfacial electron-transfer resistance (R(et)) of the electrodes were measured using an Fe(CN)(6)(3-/4-) redox probe by electrochemical impedance spectroscopy. The results showed that only a complementary sequence could form a dsDNA-PAMAM with the DNA-PAMAM probe and give an obviously enlarged R(et) value. The non-complementary and three-base mismatched sequence exhibited negligible impedance change compared with the blank measurement (the blank measurement means: ssDNA probe-modified gold electrode was directly measured by EIS). The unique spherical structure combining with more negative charges on the G-4.5 PAMAM periphery anchored on the hybrids could significantly amplify the hybridization signal (R(et) value), and the detection limit for measuring the full complementary sequence is down to pM level.