Lee B L, Chua L H, Ong H Y, Yang H G, Wu J, Ong C N
Department of Community Medicine, National University of Singapore, Kent Ridge.
Clin Chem. 1996 Sep;42(9):1405-11.
We describe a simple and sensitive HPLC method for quantifying aluminum (Al) in biological fluids by measuring the fluorescence of the Al-lumogallion complex (excitation wavelength 500 nm, emission wavelength 575 nm). Serum samples are deproteinized with 0.83 mol/L perchloric acid and centrifuged; the supernates are mixed with lumogallion reagent. Urine samples are pretreated with sodium hydroxide (2 mol/L) and methanol, kept for 1 h at -20 degrees C, and then centrifuged; the precipitate is resuspended in perchloric acid and mixed with lumogallion reagent, as for serum. The maximal fluorescence complex is formed after 1 h at pH 5 +/- 0.5. The HPLC mobile phase consists of (per liter) 100 mL, of 0.2 mol/L potassium hydrogen phthalate, 220 mL of acetonitrile, and distilled deionized water. The flow rate is 1 mL/min, and the injection volume is 5 microliters. The major aluminum species is eluted at 3.5 min, the lowest detection limit being 0.45 pg. We validated the method with samples collected from normal subjects and from workers occupationally exposed to aluminum. Comparing the results with those by traditional atomic absorption spectrometry of urinary aluminum suggests that the proposed method is reliable.
我们描述了一种简单且灵敏的高效液相色谱法，通过测量铝 - 荧光镓络合物的荧光（激发波长500 nm，发射波长575 nm）来定量生物流体中的铝。血清样品用0.83 mol/L高氯酸脱蛋白并离心；上清液与荧光镓试剂混合。尿液样品先用氢氧化钠（2 mol/L）和甲醇预处理，在-20℃下保存1小时，然后离心；沉淀物用高氯酸重悬并与荧光镓试剂混合，如同血清样品处理。在pH 5±0.5条件下1小时后形成最大荧光络合物。高效液相色谱流动相由（每升）100 mL 0.2 mol/L邻苯二甲酸氢钾、220 mL乙腈和去离子蒸馏水组成。流速为1 mL/min，进样体积为5微升。主要铝物种在3.5分钟时洗脱，最低检测限为0.45 pg。我们用从正常受试者和职业性接触铝的工人收集的样品验证了该方法。将结果与尿铝传统原子吸收光谱法的结果进行比较表明，所提出的方法是可靠的。
