Micellar electrokinetic chromatography of the anti-human immunodeficiency virus (HIV) agent michellamine B with absorption and laser-induced fluorescence detection.
Micellar electrokinetic chromatography (MEKC) was applied to the separation of the anti-HIV agents, michellamines A and B, and two other structurally related monomers found in the extract of the Ancistrocladus plants. Using buffers containing either 10 mM sodium phosphate (pH 7.0), 50 mM sodium deoxycholate and 10-20% acetonitrile or 5 mM sodium phosphate (pH 7.0), 20 mM sodium dodecyl sulfate and 25% acetonitrile allowed baseline separations of the four components in the mixture in less than 10 min. The MEKC methods gave sharper peaks and better resolution compared to high-performance liquid chromatography. For MEKC separation of the plant extracts, UV absorption detection provided adequate sensitivity; however, higher sensitivity could be achieved with UV laser-induced fluorescence detection (LIF). Using the sodium dodecyl sulfate-containing buffer and LIF, the limit of detection for michellamine B was approximately 2 ng/mL. The sensitivity was degraded approximately 100-fold when using the deoxycholate buffer because of high background fluorescence. Preliminary results show that MEKC with LIF is feasible for the sensitive detection of michellamine B in serum.
Chan
K C
KC
Program Resources, Inc./DynCorp, National Cancer Institute, Frederick Cancer Research and Development Center, MD 21702.
Majadly
F
F
McCloud
T G
TG
Muschik
G M
GM
Issaq
H J
HJ
Snader
K M
KM
D000998:Antiviral Agents / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D004586:Electrophoresis / Q000379:methods*; D015497:HIV-1 / Q000187:drug effects; D015498:HIV-2 / Q000187:drug effects; D006801:Humans; D007546:Isoquinolines / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D008823:Micelles; D015394:Molecular Structure; D009281:Naphthalenes / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D000998:Antiviral Agents; D007546:Isoquinolines; D008823:Micelles; D009281:Naphthalenes; D010936:Plant Extracts; C072235:michellamine A