Journal
FRONTIERS OF CHEMICAL SCIENCE AND ENGINEERING
Volume 7, Issue 3, Pages 322-328Publisher
HIGHER EDUCATION PRESS
DOI: 10.1007/s11705-013-1348-z
Keywords
tobramycin; aminoglycoside; refractive index detector; high-performance liquid chromatography; ionpairing
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A simple and direct method without a derivation step for routine analysis of tobramycin has been developed. This method used reversed-phase ion-pair high performance liquid chromatography (HPLC) with a refractive index (RI) detector and a C18 column which is stable at pH above 1.00. The presence of 4.50 mg . mL(-1) trifluoroacetic acid (TFA) in the mobile phase improved the protonation of tobramycin and the formation of ionpairs, and thus reduced its hydrophility. This unique separation-detection combination showed good linearity with correlation coefficients 0.9996 in the concentration range of 0.25-2.50 mg . mL(-1). The quantitation limit and detection limit were determined to be 0.23 mg . mL(-1) and 0.071 mg . mL(-1), respectively. Tobramycin was recovered in 98.00%, 98.84% and 99.64% for tobramycin solutions at concentrations of 2.25 mg . mL(-1), 1.50 mg . mL(-1) and 0.75 mg . mL(-1), respectively. The relative standard deviations for six spiked samples ranged from 0.20% to 2.40%, indicating a good reproducibility of this method.
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