title: Isolation, cytotoxic evaluation, and simultaneous quantification of eight bioactive secondary metabolites from Cicer microphyllum by high-performance thin-layer chromatography.
abstract: Chemical investigation of Cicer microphyllum resulted in the isolation and characterization of eight natural products viz. Stigmasterol, Oleanolic acid-3-acetate, Oleanolic acid, Biochanin A, Genistein, Pratensein, Chrysoeriol, and Luteolin. Herein, we report a novel, accurate, and cost-effective high-performance thin-layer chromatography method for the simultaneous quantification of the isolated natural products on silica-gel 60F254 plates using the solvent system n-hexane/ethyl acetate/formic acid (9.0:6.5:0.8, v/v/v). Natural products were quantified after postchromatographic derivatization with ceric ammonium sulfate. The method was validated as per the International Conference on Harmonization guidelines. All calibration curves showed a good linear relationship (r > 0.9943) within the test range. Precision was assessed by intra- and interday tests with relative standard deviations <1.82%, accuracy validation recovery 98.38-99.57% with relative standard deviations <1.00%. On quantification, Pratensein was a major constituent (0.921%). The screening for cytotoxic activity using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay resulted into identification of Luteolin as potent molecule with IC50 3.5 and 25.6 μg/mL against murine melanoma and human epidermoid carcinoma cell lines, respectively.
authors.affiliation: CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM), Jammu Tawi, India.
authors.forename: Alamgir A
authors.initials: AA
authors.lastname: Dar
authors.affiliation: CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM), Jammu Tawi, India.|Academy of Scientific and Innovative Research (AcSIR), CSIR-IIIM Campus, Jammu, India.
authors.forename: Santosh K
authors.initials: SK
authors.lastname: Rath
authors.affiliation: CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM), Jammu Tawi, India.
authors.forename: Afnan
authors.initials: A
authors.lastname: Qaudri
authors.affiliation: CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM), Jammu Tawi, India.
authors.forename: Buddh
authors.initials: B
authors.lastname: Singh
authors.affiliation: CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM), Jammu Tawi, India.
authors.forename: Sheikh A
authors.initials: SA
authors.lastname: Tasduq
authors.affiliation: College of Sciences, Shri Mata Vaishno Devi University, Katra, Jammu, India.
authors.forename: Anil
authors.initials: A
authors.lastname: Kumar
authors.affiliation: CSIR-Indian Institute of Integrative Medicine (CSIR-IIIM), Jammu Tawi, India.|Academy of Scientific and Innovative Research (AcSIR), CSIR-IIIM Campus, Jammu, India.
authors.forename: Payare L
authors.initials: PL
authors.lastname: Sangwan
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002623:Chemistry Techniques, Analytical / Q000191:economics / Q000379:methods* / Q000592:standards; D002855:Chromatography, Thin Layer / Q000191:economics / Q000592:standards; D029863:Cicer / Q000737:chemistry*; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D015394:Molecular Structure; D010936:Plant Extracts / Q000032:analysis / Q000302:isolation & purification / Q000633:toxicity; D015203:Reproducibility of Results
chemical_list: D010936:Plant Extracts
keywords: Cicer microphyllum; Cytotoxicity; Validation