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Isolation and Characterization of Anti-Viral Nucleoside Compounds from the Chinese Herb Taraxacum Mongolicum

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Sample Preparation

10 g dried Taraxacum mongolicum boiled in 100 mL methanol for 45 minutes

Liquid subsequently filtered with Whatman grade filter paper

Methanol roto-evaporated, leaving residue

Residue reconstituted in mobile phase to place on LC/MS LC Conditions Column: YMC C18, 2.1x250 mm, 5 μm

Flow rate: 300 μl/min

Mobile phase: A-acetonitrile, B-2 mM ammonium acetate

Gradient: 5-60% ACN in 10 min and hold for 2 min

Injection: 10 μl

Autosampler: PE series 200 MS Conditions

API 2000TM LC/MS/MS System triple quadruple mass spectrometer

Ion source: TurboIonSpray source, positive & negative modes

Scan modes: Product and precursor scans

Study Design

There are three major chemical analogs that have anti-viral activities: protease inhibitors, nucleotide, and nucleoside analogs. Protease inhibitors are synthetic compounds while nucleotide and nucleoside analogs are found in natural products. To screen nucleotides: Four nucleotide standards in adequate solutions were infused into MS/MS for optimization of the instrument. The commonproduct ions of 159 and 79 m/z were used for the precursor scans of potential nucleotide analogs in the crude extract samples under the same LC/MS/MS conditions for nucleotide standards. To screen nucleosides: Three nucleoside standards were tested under optimized MS/MS conditions. The most abundant product ion was the base fragment for all three nucleoside standards. Known nucleoside structures were searched and all base fragments were calculated. These base fragments were used for precursor ion scans of potential nucleoside analogs in the crude extract under optimized LC/MS/MS conditions for nucleoside standards.


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