Purification and Characterization of the Principal Antimutagenic Bioactive as Ethoxy-Substituted Phylloquinone from Spinach (Spinacea oleracea L.) Based on Evaluation in Models Including Human Lymphoblast TK+/-Cells.
During in vitro analysis, spinach (Spinacea oleracea L.) leaf extracts displayed varying antimutagenicity when analyzed in models including human lymphoblast (TK+/-) cell line (thymidine kinase gene mutation assay) and Escherichia coli MG1655 (rifampicin resistance assay) against chemically (ethyl methanesulfonate and 5-azacytidine) induced mutagenicity. Highest antimutagenicity was displayed by the quinone extract. The principal bioactive compound exhibited fluorescence in TLC at 366 nm (termed C4) resolved at Rf0.32 and tR15.2 min in TLC and HPLC, respectively. On the TLC plate, three spots (C1-C3), observed at 254 nm, displayed comparatively lesser antimutagenicity. Furthermore, biochemical and spectroscopic analyses using MALDI-TOF MS and NMR indicated the nature of the potent compound (C4) as an ethoxy-substituted phylloquinone derivative [2-ethoxy-3-((E)-3,7,11,15-tetramethylhexadec-2-enyl)naphthalene-1,4-dione]. The C4 compound did not display any cytotoxicity and hence possesses significant nutraceutical-based intervention possibility to combat the onset of mutation-associated disease(s).
Food Technology Division, Bhabha Atomic Research Centre , Mumbai 400 085, India.
Sanjeev
S
Kumar
Food Technology Division, Bhabha Atomic Research Centre , Mumbai 400 085, India.
Suchandra
S
Chatterjee
Food Technology Division, Bhabha Atomic Research Centre , Mumbai 400 085, India.
Jyoti
J
Tripathi
Food Technology Division, Bhabha Atomic Research Centre , Mumbai 400 085, India.
Satyendra
S
Gautam
D016587:Antimutagenic Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D002460:Cell Line; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D006801:Humans; D008214:Lymphocytes; D015394:Molecular Structure; D009152:Mutagenicity Tests; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D018724:Spinacia oleracea / Q000737:chemistry*; D010837:Vitamin K 1 / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*
D016587:Antimutagenic Agents; D010936:Plant Extracts; D010837:Vitamin K 1
nuclear magnetic resonance; prophylactic; quinone; rifampicin resistance assay; thymidine kinase assay