Development of liquid chromatographic method for the analysis of kanamycin residues in varicella vaccine using phenylisocyanate as a derivatization reagent.
A liquid chromatographic method for the determination of the aminoglycoside kanamycin in varicella vaccine is described. Kanamycin sulfate was derived with phenylisocyanate (PIC) and triethylamine for 10 min at 70 degrees C and chromatographed on a alkylamide-bonded column, Suplex pKb-100. A derivative of kanamycin sulfate was attached to four phenylisocynato groups and that molecular mass was confirmed with liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). The kanamycin-PIC derivative was found to have a retention time of 11.7 min using an eluent composed of 40% acetonitrile in water at 1.2 ml/min column flow-rate. Detection was at a wavelength of 240 nm. Recoveries ranging from 97.5 to 99.8% were found. The correlation coefficient was greater than 0.9998 over the range between 10 and 100 microg/ml. The method precision of within-day assay showed a 0.5 to 4.0% coefficient of variation (n = 5) ranging from 10 to 70 microg/ml of kanamycin concentration levels. Kanamycin-PIC derivative in reaction solution was stable for 24 h at room temperature. A simple and efficient method for the analysis of the kanamycin in varicella vaccine was developed and validated.
Analytical R&D Center, LG Chemical Ltd., Yusong-gu, Taejon, South Korea. bhkime@lgchem.co.kr
B H
BH
Kim
Y K
YK
Kim
J H
JH
Ok
D000900:Anti-Bacterial Agents / Q000032:analysis*; D019433:Chickenpox Vaccine / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004350:Drug Residues / Q000032:analysis*; D007202:Indicators and Reagents / Q000737:chemistry*; D017953:Isocyanates / Q000737:chemistry*; D007612:Kanamycin / Q000032:analysis*; D013058:Mass Spectrometry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000900:Anti-Bacterial Agents; D019433:Chickenpox Vaccine; D007202:Indicators and Reagents; D017953:Isocyanates; C025319:phenyl isocyanate; D007612:Kanamycin