A simple and precise methods for the determination of trace amounts of Cu, Fe, and Zn in lymph were investigated by graphite furnace and "one-drop method" in flame-atomic absorption spectrophotometry (GF-AAS and OD-AAS). Lymph samples were collected from cutaneous leg lymphatic vessels of rabbits by cannulation, and diluted to appropriate concentration of each metal with 0.1N-HNO
3 prior to measurements. Aliquots of 20μl for Cu and 100 μl for Fe and Zn determination were then analyzed by GF-AAS (Cu) and OD-AAS (Fe and Zn). For example, the optimum operating conditions of GF-AAS were as follows: wavelength: 324.8 nm; drying: 150 °C, ramp time: 40s, hold time: 20s; ashing I: 500°C, ramp time: 20s, hold time: 20s; ashing II: 950°C, ramp time: 15s, hold time: 20s; atomization: 2500 °C, hold time: 6s; cleaning: 2600°C, ramp time: 1s, hold time: 3s. The calibration curves for Cu, Fe, and Zn were linear up to 60 ppb, 3.0 ppm, and 0.8 ppm respectively. The detection limit (
S/
N = 3) was 2 pg for Cu, 7 ng for Fe, and 3 ng for Zn. The recovery of Cu added to lymph samples averaged (98 ± 3) % with a relative standard deviation (
n =10) of 1.0 % for 1.0 ng of Cu. The both GF-AAS and OD-AAS methods were applied to the determination of Cu, Fe, and Zn in lymph samples. As the results, the average levels of lymph Cu, Fe, and Zn in 24 healthy rabbits were (0.16
6±0.09
9) (SD)ppm, (0.91
3±0.53
6) (SD) ppm, and (1.45±0.78
1) (SD) ppm, respectively.
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