Determination of isepamicin in human plasma by HPLC with fluorescence detection after derivatization using 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate.
A simple and rapid high-performance liquid chromatographic method has been developed for determination in human plasma of isepamicin (ISP), an aminoglycoside antibiotic agent. After protein precipitation and clean-up procedure to remove lipophilic contaminants, ISP is derivatized pre-column with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate for fluorescence detection. Chromatographic separations are achieved using C(18) column and mobile phase consisting of 20 mM KH(2)PO(4) containing 8 mM triethylamine (pH 7.0) and acetonitrile (78/22, v/v). Amikacin was used as an internal standard. The calibration curve was linear over a concentration range of 0.5-50 microg/ml. The limit of quantification was 0.5 microg/ml. The intra- and inter-day variabilities of ISP were both less than 17.5%. Both derivatives were stable for at least a week at ambient condition. This assay procedure should have useful application in therapeutic drug monitoring of ISP.
Hosokawa
Shin
S
Department of Pharmacy, The Gunma University Hospital, Gunma University School of Medicine, 3-39-22 Showamachi, Maebashi, Gunma 371-8511, Japan.
Nakamura
Katsunori
K
Fujita
Yukiyoshi
Y
Horiuchi
Ryuya
R
Yamamoto
Koujirou
K
D000583:Amikacin / Q000032:analysis; D000634:Aminoquinolines / Q000737:chemistry*; D000900:Anti-Bacterial Agents / Q000097:blood*; D002219:Carbamates / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D005839:Gentamicins / Q000097:blood; D006801:Humans; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence
C078178:6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; D000634:Aminoquinolines; D000900:Anti-Bacterial Agents; D002219:Carbamates; D005839:Gentamicins; D007202:Indicators and Reagents; D000583:Amikacin; C016927:isepamicin