The Application of Trichloroacetic Acid as an Ion Pairing Reagent in LC-MS-MS Method Development for Highly Polar Aminoglycoside Compounds
2010
Cheng, Chang | Liu, Shaorong | Xiao, Deqing | Hansel, Steven
A simple, sensitive and robust liquid chromatography-tandem mass spectrometry method was developed and validated for highly polar aminoglycoside compounds gentamicin, kanamycin and apramycin. The effect of trichloroacetic acid (TCA) concentration on plasma protein precipitation and sample recovery was studied and an optimized concentration of 25-30% TCA were determined that gives the best sample recovery for aminoglycosides from rat plasma. The effect of TCA concentration on the chromatographic behavior of gentamicin and tobramycin was studied on a Synergy Max RP-column using a mobile phase with a pH of 2.78. Other than protein precipitation, TCA also acted as ion pairing reagent and was only present in the samples but not in the mobile phases. The data demonstrated that by increasing the TCA concentration, the analyte retention and sensitivity were improved. The absence of TCA in mobile phase helped to reduce the ion source contamination and to achieve good reproducibility. The plasma method was linearly calibrated from 1-5,000, 20-10,000, 10-10,000 ng mL⁻¹ with precisions of 2.6-4.1, 3.3-5.0, 1.5-9.9%, and accuracies of 94.7-103.7, 87.9-104.9, 91.3-103.6% for gentamicin, kanamycin and apramycin, respectively. The LLOQs corresponding with a coefficient of variation less than 20% were 1, 20 and 10 ng mL⁻¹ for gentamicin, kanamycin and apramycin, respectively.
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