Bioassay-Guided Fractionation of the Active Constituent of Juniperus Chinensis
Deoxypodophyllotoxin, a lignan, was afforded from the bioassay-guided fractionation of the EtOAc soluble part of the leaves and twigs of Juniperus chinensis. The fractionation was directed by microtitration cytotoxicity assay employing human cervical adenocarcinoma (HeLa) cell line. The activity...
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| Format: | Thesis |
| Language: | English English |
| Published: |
1997
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| Online Access: | http://psasir.upm.edu.my/id/eprint/8382/1/FSMB_1997_11_A.pdf http://psasir.upm.edu.my/id/eprint/8382/ |
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| Summary: | Deoxypodophyllotoxin, a lignan, was afforded from the bioassay-guided
fractionation of the EtOAc soluble part of the leaves and twigs of Juniperus
chinensis. The fractionation was directed by microtitration cytotoxicity assay
employing human cervical adenocarcinoma (HeLa) cell line. The activity was visible
by fixing and staining the cells and comparing the number of cell reduction by the
active agent with the confluent controls. A judicious combination of chromatographic
techniques was adopted in purifying the active compound from the crude complex.
The structure of the isolated lignan was elucidated using spectroscopic techniques
including ultraviolet spectroscopy (UV), infrared spectroscopy (IR), nuclear magnetic, resonance spectroscopy (¹H and ¹³C-NMR), mass spectroscopy (MS), and also by
comparison with the literature.The cytotoxic concentration of deoxypodophyllotoxin which caused up to
almost 100% reduction of HeLa cells was determined as 0.004 Jlg/ml. Cytotoxic
activity of this lignan was further evaluated on different types of specific human organ
tumour cell lines: KU8 12F (Chronic mylogeneous leukemia), TK-10 (Renal
carcinoma), UACC-62 (Melanoma) as well as CEM-SS (T-cell lymphoblastic
leukemia). All of the tumour cell lines studied were found to be susceptible to
deoxypodophyllotoxin, nevertheless, the degree of susceptibilities was different
between cell lines. Minimum effective concentration (MEC) with almost 1 00%
reduction of the cells were observed in HeLa (0.004 µg/ml), TK-10 (0.01 µg/ml),
UACC-62 (0.004 µg/ml) and CEM-SS (0.01 µg/ml). Whilst KU8 12F (0.04 µg/ml)
inhibited only 50% the cell growth (ECso). Thus, the most sensitive cell l ines towards
the treatment of the lignan were HeLa and UACC-62.
Antimicrobial disc diffusion assay (Bauer et al., 1966) on
deoxypodophyllotoxin was carried out employing gram positive bacteria (Bacillus
megaterium, Bacillus cereus, Bacillus subtilis, Flavobacterium meningosepticum,
Slaphylloccus aureus, Micrococcus luleus, Chrysomollas leuteola and Aeromonas
salmonella), and gram negative bacteria (Pseudomonas aeruginosa, Pseudomonas
paucinobilis, Pseudomonas capacia and Escherichia coli), and on yeast (Torulopsis glabrata, Crytococcus neoformans, Saccharomyces lipolytica, Candida albicans,
Candida lipolytica and Candida inlermedia), and also a fungi (Aspergillus
ochraceous). The growth of most of the organisms were inhibited by deoxypodophyllotoxin at the concentration of 10 mg/ml by producing a clearing zone
with diameter ranging between 8 to 12 mm with the exception of Pseudomonas
aeruginosa, P. paucinobilis, Aeromonas salmonella and Candida intermedia. |
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