Test Performance Criteria / Testing Method
System Suitability Column efficiency of >1200
Five replicate injections RSDa ≤ 2.0%
Asymmetry < 1.5
Resolution >1.25
Capacity Factor (C1) between 2 and 7
Robustness ±2.5% change in methanol composition in mobile phase, ±5°C change in derivatization temperature, ±10% change of mobile phase flow rate, different injection volume (10, 15, and 20 µl)
Specificity No interfering peaks of the same retention time as either Gentamicin Sulfate or L-leucine with peak area above 1% of the corresponding lowest standard for both non stressed and acid stressed samples
Linearity R2> 0.98
Accuracy/Recovery 85 – 115% of Theoretical (80 – 120% at LOQb)
Repeatability ≤ 15% RSDa(≤ 20% RSDa at LOQb)
Intermediate Precision ≤ 15% RSDa(≤ 20% RSDa at LOQb)
LOQb Tested at: Gentamicin Sulfate = 50 µg/mL
L-leucine = 0.5 µg/mL
LODc Tested at: Gentamicin Sulfate = 5 µg/mL
L-leucine = 0.2 µg/mL
aRelative Standard Deviation
bLimits of Quantification
c Limits of Detection
Table 2: Analytical qualification performance criteria for the determination of Gentamicin Sulfate and L-leucine in formulations.