Chen Hua, Gong Zhengjun, Zhang Zhujun
College of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044, China.
J Pharm Biomed Anal. 2006 Jun 16;41(4):1412-7. doi: 10.1016/j.jpba.2006.02.050. Epub 2006 Apr 17.
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.
采用在线微透析采样与流动注射化学发光检测（FI-MD-CL）联用系统研究了甲硝唑（MTZ）与人血清白蛋白（HSA）的相互作用。将感兴趣的药物和HSA以不同摩尔比在0.067 mol L(-1)磷酸盐缓冲液（pH 7.4）中混合，并在37℃水浴中孵育。然后将微透析探针放入MTZ-HSA混合溶液中，以5 μL min(-1)的灌注速率进行采样。采用流动注射化学发光法测定微透析液。实验条件下MTZ的体外回收率（R）约为25.2%，相对标准偏差约为3.2%。结合常数（K）和结合位点数（n）的估计值分别为1.50×10(3)L mol(-1)和1.89。用Scatchard分析和Klotz图得到的nK值非常相似。该方法为研究药物-蛋白质相互作用提供了一种可靠且简便的技术。
