Coupling microdialysis with flow-injection chemiluminescence detection for a protein-drug interaction study.
The interaction of metronidazole (MTZ) and human serum albumin (HSA) was studied using the coupling system of on-line microdialysis sampling with flow-injection chemiluminescence detection (FI-MD-CL). The interested drug and HSA were mixed in different molar ratios in 0.067 mol L(-1) phosphate buffer (pH 7.4) and incubated at 37 degrees C in a water-bath. Then the microdialysis probe was put into the MTZ-HSA mixed solution and sampled at a perfusion rate of 5 microL min(-1). The microdialysates was determined using flow-injection chemiluminescence. In vitro recovery (R) of MTZ under experimental conditions was approximately 25.2% with a R.S.D. of about 3.2%. The values estimated for the binding constant (K) and the number of the binding sites (n) were found to be 1.50 x 10(3)L mol(-1) and 1.89, respectively. The values of nK obtained using Scatchard analysis and Klotz plot were found to be quite similar. The method provided a reliable and simple technique for the study of drug-protein interaction.
Chen
Hua
H
College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044, China. chenhuacqu@yahoo.com.cn
Gong
Zhengjun
Z
Zhang
Zhujun
Z
D000890:Anti-Infective Agents / Q000378:metabolism*; D004347:Drug Interactions; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods; D008795:Metronidazole / Q000378:metabolism*; D017551:Microdialysis; D012709:Serum Albumin / Q000378:metabolism*
D000890:Anti-Infective Agents; D012709:Serum Albumin; D008795:Metronidazole