Table 2

Freezer stability data for human and mouse hepcidin

Nominal hepcidin standard addition, ng/mLAverage calculated concentration, ng/mLStandard deviation, ng/mL% RSD% REn
Human serum       
    Nonspiked N/A <1.0 N/A N/A N/A 6 
        3× freeze-thaw nonspiked N/A <1.0 N/A N/A N/A 6 
        Seven-wk freezer stability nonspiked N/A <1.0 N/A N/A N/A 6 
    Spiked 5 4.6 0.74 16.0 −7.3 6 
        3× freeze-thaw spiked 5 4.6 0.22 4.8 −7.7 6 
        Seven-wk freezer stability spiked 5 4.18 0.40 9.5 −16.4 6 
    Spiked 50 59.4 4.23 7.1 18.9 6 
        3× freeze-thaw spiked 50 57.8 4.80 8.3 15.6 6 
        Seven-wk freezer stability spiked 50 44.0 6.23 14.2 −12.1 6 
    Dilutional linearity: 100 × dilution† 5000 5090.0 6.00 11.8 1.7 6 
Mouse serum       
    Nonspiked N/A 3.15* 0.48 15.2 N/A 6 
        3× freeze-thaw nonspiked N/A 3.12 0.20 6.6 −1.1 6 
        Eight-wk freezer stability nonspiked N/A 2.49 0.23 0.09 −20.7 6 
    Spiked 20 21.10 1.69 8.0 −8.9 6 
        3× freeze-thaw spiked 20 19.89 0.96 4.8 −14.1 6 
        Eight-wk freezer stability spiked 20 16.90 1.57 9.3 −24.9 6 
    Spiked 200 159.15 13.60 8.5 −21.7 6 
        3× freeze-thaw spiked 200 149.31 5.63 3.8 −26.5 6 
        Eight-wk freezer stability spiked 200 183.74 8.08 4.4 −9.3 6 
    Dilutional linearity: 100 × dilution† 5000 4114.0 5.34 13.0 −17.8 6 
Nominal hepcidin standard addition, ng/mLAverage calculated concentration, ng/mLStandard deviation, ng/mL% RSD% REn
Human serum       
    Nonspiked N/A <1.0 N/A N/A N/A 6 
        3× freeze-thaw nonspiked N/A <1.0 N/A N/A N/A 6 
        Seven-wk freezer stability nonspiked N/A <1.0 N/A N/A N/A 6 
    Spiked 5 4.6 0.74 16.0 −7.3 6 
        3× freeze-thaw spiked 5 4.6 0.22 4.8 −7.7 6 
        Seven-wk freezer stability spiked 5 4.18 0.40 9.5 −16.4 6 
    Spiked 50 59.4 4.23 7.1 18.9 6 
        3× freeze-thaw spiked 50 57.8 4.80 8.3 15.6 6 
        Seven-wk freezer stability spiked 50 44.0 6.23 14.2 −12.1 6 
    Dilutional linearity: 100 × dilution† 5000 5090.0 6.00 11.8 1.7 6 
Mouse serum       
    Nonspiked N/A 3.15* 0.48 15.2 N/A 6 
        3× freeze-thaw nonspiked N/A 3.12 0.20 6.6 −1.1 6 
        Eight-wk freezer stability nonspiked N/A 2.49 0.23 0.09 −20.7 6 
    Spiked 20 21.10 1.69 8.0 −8.9 6 
        3× freeze-thaw spiked 20 19.89 0.96 4.8 −14.1 6 
        Eight-wk freezer stability spiked 20 16.90 1.57 9.3 −24.9 6 
    Spiked 200 159.15 13.60 8.5 −21.7 6 
        3× freeze-thaw spiked 200 149.31 5.63 3.8 −26.5 6 
        Eight-wk freezer stability spiked 200 183.74 8.08 4.4 −9.3 6 
    Dilutional linearity: 100 × dilution† 5000 4114.0 5.34 13.0 −17.8 6 

N/A indicates not applicable.

*

Concentration was determined by analyzing 6 replicates without standard addition.

†

The 5000 ng hepcidin/mL spiked samples were diluted 100× in control rabbit serum to yield a nominal concentration of 50 ng/mL. The values shown are corrected for the dilution factor

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