Kawasaki N, Tanimoto T, Tanaka A, Hayakawa T, Miyasaka N
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo, Japan.
J Chromatogr B Biomed Appl. 1994 Jun 17;656(2):436-40. doi: 10.1016/0378-4347(94)00135-9.
Non-protein-bound iron in human synovial fluid was determined using high-performance liquid chromatography with electrochemical detection. The procedure was based on the separation of the iron-diethylenetriaminepentaacetic acid (DTPA) complex formed directly on a chromatographic column containing an anion-exchange resin followed by electrochemical detection. The method enabled more than 0.1 microM Fe(III) to be determined with an injection volume of 10 microliters. A mixture of synovial fluid, 20 microM DTPA and acetate buffer was incubated in the presence and absence of superoxide (O2-) generated by a xanthine-xanthine oxidase system and was ultrafiltered through a 30,000 molecular mass cut-off filter. No iron was detected in the ultrafiltrate at physiological pH. However, the presence of iron was observed in the ultrafiltrate at low pH, and O2- facilitated the release of iron into the synovial fluid. This result suggested that in an inflamed joint with generated O2- and decreased pH, iron may be released into the synovial fluid.
采用高效液相色谱-电化学检测法测定人滑液中的非蛋白结合铁。该方法基于在含有阴离子交换树脂的色谱柱上直接形成的铁-二乙烯三胺五乙酸（DTPA）络合物的分离，随后进行电化学检测。该方法能够在进样体积为10微升的情况下测定超过0.1微摩尔的Fe(III)。将滑液、20微摩尔DTPA和乙酸盐缓冲液的混合物在存在和不存在由黄嘌呤-黄嘌呤氧化酶系统产生的超氧化物（O2-）的情况下孵育，并通过截留分子量为30,000的滤膜进行超滤。在生理pH值下，超滤液中未检测到铁。然而，在低pH值下，超滤液中观察到铁的存在，并且O2-促进了铁释放到滑液中。该结果表明，在产生O2-且pH值降低的炎症关节中，铁可能会释放到滑液中。
