Isolation, characterisation, antifungal activity and validated UPLC/MS/MS method for quantification of novel compound from Artemisia tournefortiana Reichb.

Investigation into the chemical diversity of Artemisia tournefortiana resulted in isolation of one novel compound named tournefortin A and two known artemetin and tournefortin B bioactive compounds. Tournefortin B is first time obtained from natural source. The structure of all the isolated compounds were elucidated by detailed 1D and 2D NMR including HSQC, HMBC, 1H-1HOSY and NOESY spectroscopic techniques. Minimum inhibitory concentration (MIC) of all the tested compounds against tested fungal strains lies between 0.4 and 6.4 µg/mL and lowest MIC of 0.4 µg/mL of compound tournefortin A was found against Alternaria alternate. All the isolated compounds were quantified through UPLC/MS/MS and the developed method will serve as a first fingerprint method for the rapid determination of these phytomolecules in various plant extracts. Tournefortin B was found to be present in higher concentration. The higher antifungal effect of the isolated compounds suggests that this plant could act as potential source of antimicrobial agents.[Formula: see text].

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

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.