Pre-column derivatization of sisomicin with o-phthalaldehyde-beta-mercaptopropionic acid and its application to sensitive high-performance liquid chromatographic determination with fluorimetric detection.
The stability of the o-phthalaldehyde (OPA) derivatives of sisomicin obtained using beta-mercaptopropionic acid was investigated by reversed-phase high-performance liquid chromatography. One of the fluorescent derivatives of sisomicin was stable at least for 6 h in 50% methanol under the optimal conditions used (OPA concentration, pH and temperature). When plasma samples spiked with sisomicin were analysed, the response was linear in the calibration range 136-900 pg of sisomicin per injected volume (40 microliters). As little as 0.06 micrograms of sisomicin per 1 ml of plasma could be detected with signal-to-noise ratio greater than or equal to 2. For plasma samples spiked with 0.2 micrograms/ml sisomicin, the recovery was 97.1 +/- 6.6% (mean +/- S.D., n = 5) with a within-run coefficient of variation of 6.8% and a day-to-day coefficient of variation of 7.2%. The method was also applied to plasma samples from rabbit after a subcutaneous injection of 1 mg/kg sisomicin.
Tawa
R
R
Department of Analytical Chemistry, Kyoto Pharmaceutical University, Japan.
Koshide
K
K
Hirose
S
S
Fujimoto
T
T
D015097:3-Mercaptopropionic Acid; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007273:Injections, Intramuscular; D011817:Rabbits; D012853:Sisomicin / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D013050:Spectrometry, Fluorescence; D009764:o-Phthalaldehyde
D009764:o-Phthalaldehyde; D015097:3-Mercaptopropionic Acid; D012853:Sisomicin