Document 0184
 DOCN  M9460184
 TI    Determination of michellamine B in biological fluids by high-performance
       liquid chromatography with fluorescence detection.
 DT    9404
 AU    Supko JG; Malspeis L; Laboratory of Pharmaceutical Chemistry, National
       Cancer; Institute, Frederick, Maryland 21701-8527.
 SO    Anal Biochem. 1994 Jan;216(1):52-60. Unique Identifier : AIDSLINE
       MED/94182739
 AB    The National Cancer Institute is pursuing preclinical development of
       michellamine B (MB), a novel dimeric polyhydroxylated
       naphthalene-tetrahydroisoquinoline alkaloid isolated from Ancistrocladus
       abbreviatus, as an anti-human immunodeficiency virus (HIV) agent. MB
       protects human lymphoid cells from the cytopathic effects of both HIV-1
       and HIV-2 in vitro. A specific, sensitive, and convenient method for
       assaying the compound in biological fluids has been developed. Samples
       were prepared for analysis by initial treatment with dilute
       trichloroacetic acid followed by thorough mixing with a solution of the
       internal standard (alpha-naphthoflavone) in acetonitrile to denature
       macromolecules. The supernatant afforded by centrifugation, upon
       dilution with the aqueous component of the liquid chromatographic
       eluent, was loaded onto a 4-microns Nova-Pak phenyl column (3.9 mm x 15
       cm). Chromatography was performed at ambient temperature using an
       isocratic mobile phase composed of 10 mM octyl sodium sulfate and 15
       microM tetrabutylammonium hydrogen sulfate in acetonitrile/0.05 M
       ammonium formate buffer, pH 4.0 (46/54, v/v), at a flow rate of 0.6
       ml/min. The intense native fluorescence of MB, which exhibited
       excitation and emission maxima in the mobile phase at 232 and 393 nm,
       respectively, provided a highly sensitive and selective means of
       detection. Mean values of the retention times for the drug and internal
       standard determined over 11 months were 10.71 +/- 0.53 and 13.14 +/-
       0.52 min, respectively (SD, n = 52). Employing a sample volume of 50
       microliters, the lowest concentration of MB included in the standard
       curves of mouse, dog, and human plasma, 10 ng/ml (11.4 nM), was
       quantified with coefficients of variation less than 10%.(ABSTRACT
       TRUNCATED AT 250 WORDS)
 DE    Animal  Antiviral Agents/*ANALYSIS  Chromatography, High Pressure Liquid
       Dogs  Human  Isoquinolines/*ANALYSIS/PHARMACOKINETICS  Male  Mice
       Naphthalenes/*ANALYSIS/PHARMACOKINETICS  Temperature  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).