Skip to Content
Merck
  • Simultaneous determination of a new anthracycline, DA-125, and its metabolites M1, M2, M3 and M4 in plasma and urine by high-performance liquid chromatography.

Simultaneous determination of a new anthracycline, DA-125, and its metabolites M1, M2, M3 and M4 in plasma and urine by high-performance liquid chromatography.

Journal of chromatography. B, Biomedical applications (1994-06-17)
H J Shim, E D Lee, E J Yoon, S D Lee, W B Kim, J Yang, M G Lee
ABSTRACT

A high-performance liquid chromatographic method was developed for the simultaneous determination of a new anthracycline, DA-125 (I), and its metabolites (M1, M2, M3, and M4) in rat plasma and urine using fluorescein as an internal standard. Compound I, a prodrug of M1, is a beta-alanine derivative of M1, and only M1 shows antineoplastic activity. The method involved extraction or deproteinization followed by injection of 80-100 microliters of the aqueous layer or supernatant onto a C18 reversed-phase column. The mobile phases were 1% acetic acid-isopropyl alcohol-methanol (70:20:10, v/v) or 5 mM of ion-pairing chromatography reagent (IPC B8)-isopropyl alcohol-methanol (70:20:10, v/v) for the extraction or deproteinization methods, respectively. The flow-rate was 1.5 ml/min for both methods. The column effluent was monitored by a fluorescence detector with excitation wavelength of 488 nm and emission wavelength of 556 nm. The detection limits for M1, M2, M3, and M4 in rat plasma and urine were 50 ng/ml for all compounds using the extraction method, and 100, 50, 50, 50, and 50 ng/ml for I, M1, M2, M3 and M4 in rat plasma respectively, using the deproteinization method. No interferences from endogenous substances, adriamycin or daunorubicin were found.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
4-Methyl-2-pentanone, suitable for HPLC, ≥99.5%
Supelco
4-Methyl-2-pentanone, analytical standard
Sigma-Aldrich
4-Methyl-2-pentanone, ACS reagent, ≥98.5%
Sigma-Aldrich
4-Methyl-2-pentanone, ≥99%
Sigma-Aldrich
4-Methyl-2-pentanone, ≥99%, FCC