Iron, for biosynthesis is transported through the bloodstream by the protein transferrin. The analysis for iron in transferrin requires only about 1 ug for an accuracy of 2-5%. If blood is collected without an anticoagulant, the blood clots and the liquid that remains is called serum. Serum normally contains about 1 ug of Fe/ml attached to the transferrin. Serum was analyzed in the following manner: To 1.00 ml of sample are added 2.00 ml of reducing agent and 2.00 ml of acid to reduce and release Fe from transferrin. The serum proteins are precipitated with 1.00 ml of 30% trichloracetic acid. The volume change of the solution is negligible when the protein precipitates and remains at 6.00 ml. A 4.00 ml sample of the liquid is transferred to a fresh test tube and is treated with 1.00 ml of solution containing ferrozine and buffer. The absorbance of this solution is measured after a 10 min waiting period. For an absorbance standard, a volume of 1.00 ml containing 3.00 ug Fe is diluted to 6.00 ml with other reagents. The following data were obtained
Sample Absorbance at 562 nm in 1.0 cm cell
Blank 0.038
Fe standard 0.239
Serum sample 0.129
What is the concentration of iron in the serum? Calculate the molar absorptivity of (ferrozine)3Fe 4-
Iron, for biosynthesis is transported through the bloodstream by the protein transferrin. The analysis for iron in transferrin requires only about 1 ug for an accuracy of 2-5%. If blood is collected without an anticoagulant, the blood clots and the liquid that remains is called serum. Serum normally contains about 1 ug of Fe/ml attached to the transferrin. Serum was analyzed in the following manner: To 1.00 ml of sample are added 2.00 ml of reducing agent and 2.00 ml of acid to reduce and release Fe from transferrin. The serum proteins are precipitated with 1.00 ml of 30% trichloracetic acid. The volume change of the solution is negligible when the protein precipitates and remains at 6.00 ml. A 4.00 ml sample of the liquid is transferred to a fresh test tube and is treated with 1.00 ml of solution containing ferrozine and buffer. The absorbance of this solution is measured after a 10 min waiting period. For an absorbance standard, a volume of 1.00 ml containing 3.00 ug Fe is diluted to 6.00 ml with other reagents. The following data were obtained
Sample Absorbance at 562 nm in 1.0 cm cell
Blank 0.038
Fe standard 0.239
Serum sample 0.129
What is the concentration of iron in the serum? Calculate the molar absorptivity of (ferrozine)3Fe 4-
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