2. Academic Validation
  3. Isolation, cytotoxicity evaluation and HPLC-quantification of the chemical constituents from Prangos pabularia

Isolation, cytotoxicity evaluation and HPLC-quantification of the chemical constituents from Prangos pabularia

  • PLoS One. 2014 Oct 14;9(10):e108713. doi: 10.1371/journal.pone.0108713.
Saleem Farooq 1,  Shakeel-u-Rehman 2,  Nisar Ahmad Dangroo 1,  Dev Priya 3,  Javid Ahmad Banday 4,  Pyare Lal Sangwan 1,  Mushtaq Ahmad Qurishi 4,  Surrinder Koul 1,  Ajit Kumar Saxena 3
Affiliations

Affiliations

  • 1 Bio-organic Chemistry Section, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
  • 2 Bio-organic Chemistry Section, CSIR-Indian Institute of Integrative Medicine, Srinagar, India; Department of Chemistry, University of Kashmir, Srinagar, India.
  • 3 Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
  • 4 Department of Chemistry, University of Kashmir, Srinagar, India.
PMID: 25314269 DOI: 10.1371/journal.pone.0108713
Abstract

Phytochemical analysis of the dichloromethane:methanol (1:1) extract of root parts of Prangos pabularia led to the isolation of twelve cytotoxic constituents, viz., 6-hydroxycoumarin (1), 7-hydroxycoumarin (2), heraclenol-glycoside (3), xanthotoxol (4), heraclenol (5), oxypeucedanin hydrate (6), 8-((3,3-dimethyloxiran-2-yl)methyl)-7-methoxy-2H-chromen-2-one (7), oxypeucedanin hydrate monoacetate (8), xanthotoxin (9), 4-((2-hydroxy-3-methylbut-3-en-1-yl)oxy)-7H-furo[3,2-g]chromen-7-one (10), imperatorin (11) and osthol (12). The isolates were identified using spectral techniques in the LIGHT of literature. 3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity screening of the isolated constituents was carried out against six human Cancer cell lines including lung (A549 and NCI-H322), epidermoid carcinoma (A431), melanoma (A375), prostate (PC-3) and Colon (HCT-116) cell lines. Osthol (12) exhibited the highest cytotoxicity with IC50 values of 3.2, 6.2, 10.9, 14.5, 24.8, and 30.2 µM against epidermoid carcinoma (A431), melanoma (A375), lung (NCI-H322), lung (A549), prostate (PC-3) and colon (HCT-116) cell lines respectively. Epidermoid carcinoma cell line A431 was sensitive to most of the compounds followed by lung (A549) Cancer cell line. Finally a simple and reliable HPLC method was developed (RP-HPLC-DAD) and validated for the simultaneous quantification of these cytotoxic constituents in Prangos pabularia. The extract was analyzed using a reversed-phase Agilent ZORBAX eclipse plus column C18 (4.6×250 mm, 5 µm) at 250 nm wavelength using a gradient water-methanol solvent system at a flow rate of 0.8 ml/min. The RP-HPLC method is validated in terms of recovery, linearity, accuracy and precision (intra and inter-day validation). This method, because of shorter analysis time, makes it valuable for the commercial quality control of Prangos pabularia extracts and its future pharmaceutical preparations.

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