Coupling immunoassays with chromatographic separation techniques

2000 ◽  
Vol 14 (6) ◽  
pp. 442-449 ◽  
Author(s):  
Zhe Tang ◽  
H. Thomas Karnes
Author(s):  
HAITHAM ALI IBRAHIM ◽  
FATEHIA SAYED ELSHARAWY ◽  
MAHMMOUD ELHASSAB ◽  
SAMAH SHABANA ◽  
EMAN GABER HAGGAG

Objective: phytochemical investigation of the ethyl acetate fraction (EAF) of 80% aqueous methanol extract (AME) of Dypsis leptocheilos leaves, in addition to evaluation of the antioxidant, cytotoxic and antimicrobial activities of the AME and EAF. Docking was used to predict and understand cytotoxicity of the isolated compounds. Methods: The ethyl acetate fraction (EAF) of Dypsis leptocheilos leaves was subjected to different chromatographic separation techniques. Structures of the isolated compounds were established by different spectroscopic techniques (1H/13C NMR). Antioxidant activity was evaluated by DPPH assay, while cytotoxicity was evaluated by MTT cell viability assay. Antimicrobial activity was evaluated by agar diffusion method. The docking study was conducted using Auto Dock Vina; the estrogen receptor (PDB 5t92) was used as a receptor for the docking. Results: Chromatographic separation techniques were led to the isolation of five phenolic compounds; these compounds were identified to be apigenin 8-C-β-D-glucopyranoside (Vitexin) (1), apigenin 6-C-β-D-glucopyranoside (Isovitexin) (2), luteolin 7-O-β-D-glucopyranoside (3), luteolin 8-C-β-D-glucopyranoside (Orientin) (4), luteolin 6-C-β-D-glucopyranoside (Isoorientin) (5). They were isolated and identified for the first time from this plant species. The AME and EAF showed moderate activity against Gram positive and Gram negatvie bacteria, while both of them showed similar and powerful antioxidant activity with SC50 = 12.8±0.56 µg/ml and SC50 = 17±0.77 µg/ml respectively, compared to ascorbic (reference drug) SC50 = 14.2±0.35 µg/ml. The EAF showed higher cytotoxic activity on the MCF-7 cells (human breast cancer cell line), with IC50 = 12.3 ± 1.82 µg/ml, compared to Vinblastine Sulfate (reference drug). All isolated compounds showed good binding affinity to the estrogen receptors existed in the MCF-7 cell. Conclusion: Five phenolic compounds were isolated for the first time from the EAF of Dypsis leptocheilos leaves. The AME and EAF extracts showed variable antioxidant, antimicrobial and cytotoxic activities.


1994 ◽  
Vol 349 (5) ◽  
pp. 338-345 ◽  
Author(s):  
Karl Cammann ◽  
Wolfgang Kleib�hmer ◽  
Elmar Mussenbrock ◽  
Bernd Ro� ◽  
Frank Zuther

2011 ◽  
Vol 690 (2) ◽  
pp. 148-161 ◽  
Author(s):  
Christophe Tistaert ◽  
Bieke Dejaegher ◽  
Yvan Vander Heyden

1998 ◽  
Vol 75 (10) ◽  
pp. 1209
Author(s):  
John T. Riley ◽  
Thomas K. Green ◽  
Darwin B. Dahl

1987 ◽  
Vol 14 (4) ◽  
pp. 429 ◽  
Author(s):  
PJ Hall ◽  
J Badenoch-Jones ◽  
CW Parker ◽  
DS Letham ◽  
BA Barlow

Cytokinins present in the xylem saps of two species of Amyema mistletoes and three species of Eucalyptus hosts have been measured using radioimmunoassays employing antibodies elicited against zeatin riboside, dihydrozeatin riboside and isopentenyladenosine. The radioimmunoassays, when used in combination with enzymatic treatments and chromatographic separation techniques, allowed the detection of a wide variety of cytokinin bases, ribosides, O-glucosides and nucleotides. Column chromatography over paraffin-coated silica gel was used to separate cytokinins from contaminating polar plant compounds. Results demonstrated a closer relationship between the types and concentrations of cytokinins in the xylem saps of mistletoes and their hosts in a mimicking situation (A. miquelii on E. polyanthemos) than in a non-mimicking situation (A. pendulum on E. cinerea).


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