Fluorescent reversible regulation based on photoinduced electron transfer from DNA to quantum dots and intercalation binding of DNA intercalator to DNA.

Based on the fluorescent reversible regulation, a novel sensor platform was designed for the detection of DNA intercalators utilizing the intercalation binding of DNA intercalators to DNA as an inherent exhibition and the fluorescence change of quantum dots (QDs) as an external manifestation. To prove its feasibility, acridine orange (AO) was chosen as an example of DNA intercalator. When different concentrations of herring sperm DNA (hsDNA) were added to cysteamine (CA)-capped ZnSe QDs solution, the hsDNA bound with the QDs through electrostatic interaction due to the photoinduced electron transfer from hsDNA to QDs and formed QDs-hsDNA complexes with 1:1 ratio, leading to the fluorescence quenching of the QDs; and upon addition of different concentrations of AO to the QDs-hsDNA complex system, the AO first caused the release of the hsDNA from the complexes and concomitantly bound with them through intercalation binding and formed AO-hsDNA complexes with 1:3 ratio on account of the fact that the intercalation binding constant between AO and hsDNA (1.932 × 105 L/mol) was greater than the electrostatic interaction constant between QDs and hsDNA (7.874 × 104 L/mol), resulting in the fluorescence recovery of the QDs. Therefore, the detection of AO could be achieved through the relationship between the fluorescence recovery yield of the QDs and the concentration of AO added. The results illustrated that the fluorescence recovery yield of the QDs-hsDNA system was linearly dependent to the concentration of AO in the range of 5.0-75.0 × 10-5 mol/L with a detection limit (3σ/K) of 1.5 × 10-5 mol/L. This dual-directional fluorescent regulation provided a novel method for the detection of DNA intercalators such as polycyclic aromatic hydrocarbons and drugs interfering with DNA-synthesis and possessed some potential applications in the investigation of the interactions between DNA intercalators and DNA.

Development of propidium iodide as a fluorescence probe for the on-line screening of non-specific DNA-intercalators in Fufang Banbianlian Injection.

Fufang Banbianlian Injection (FBI) has been widely used as an anti-inflammatory and anti-tumor prescription. To understand the relationships between its bioactive ingredients and pharmacological efficacies, our previous study has been successfully identified some DNA-binding compounds in FBI using an established on-line screening system, in which 4',6-diamidino-2-phenylindole (DAPI) was developed as a probe. However, DAPI can be only used to screen ATT-specific DNA minor groove binders, leaving the potential active intercalators unknown in FBI. As a continuation of our studies on FBI, here we present a sensitive analytical method for rapid identification and evaluation of DNA-intercalators using propidium iodide (PI) as a fluorescent probe. We have firstly established the technique of high-performance liquid chromatography-diode-array detector-multistage mass spectrometry-deoxyribonucleic acid-propidium iodide-fluorescence detector (HPLC-DAD-MS(n)-DNA-PI-FLD) system. As a result, 38 of 58 previously identified compounds in FBI were DNA-intercalation active. Interestingly, all previously reported DNA-binders also showed intercalative activities, suggesting they are dual-mode DNA-binders. Quantitative study showed that flavonoid glycosides and chlorogenic acids were the main active compounds in FBI, and displayed similar DNA-binding ability using either DAPI or PI. In addition, 13 active compounds were used to establish the structure-activity relationships. In this study, PI was developed into an on-line method for identifying DNA-intercalators for the first time, and thus it will be a useful high-throughput screening technique for other related samples.

Separation and determination of psoralen and isopsoralen by microemulsion electrokinetic chromatography.

The use of microemulsion electrokinetic chromatography was proposed to separate psoralen and isopsoralen in Psoralea corylitolia L. and its preparations. After conducting a series of optimizations, baseline separation was obtained for the analytes under the optimum conditions [sodium dodecyl sulfate 1.05% (m/v), ethyl acetate 0.96% (v/v), butan-1-ol 0.24% (v/v), 25 mm borate, pH 8.5, applied voltage 17.5 kV and detection at 254 nm]. Regression equations revealed linear relationships (correlation coefficients 0.9997 for psoralen and 0.9999 for isopsoralen) between the peak area of each analyte and the concentration. The limits of detection (defined as a signal-to-noise ratio of about 3) were 0.42 microg/mL for psoralen and 0.32 microg/mL for isopsoralen, respectively. The analytes were successfully determined with recoveries ranging from 95.50 to 102.03%. The method has been successfully applied for the analysis of psoralen and isopsoralen in medical samples. Furthermore, a simple and effective extraction method, with methanol in an ultrasonic water bath for 20 min three times, was used for sample preparation.