HPLC-PDA Method for Quantification of Bioactive Compounds in Crude Extract and Fractions of Aucklandia costus Falc. and Cytotoxicity Studies against Cancer Cells

Abstract

Aucklandia costus Falc. (Synonym: Saussurea costus (Falc.) Lipsch.) is a perennial herb of the family Asteraceae. The dried rhizome is an essential herb in the traditional systems of medicine in India, China and Tibet. The important pharmacological activities reported for Aucklandia costus are anticancer, hepatoprotective, antiulcer, antimicrobial, antiparasitic, antioxidant, anti-inflammatory and anti-fatigue activities. The objective of this study was the isolation and quantification of four marker compounds in the crude extract and different fractions of A. costus and the evaluation of the anticancer activity of the crude extract and its different fractions. The four marker compounds isolated from A. costus include dehydrocostus lactone, costunolide, syringin and 5-hydroxymethyl-2-furaldehyde. These four compounds were used as standard compounds for quantification. The chromatographic data showed good resolution and excellent linearity (r2 ˃ 0.993). The validation parameters, such as inter- and intraday precision (RSD < 1.96%) and analyte recovery (97.52–110.20%; RSD < 2.00%),revealed the high sensitivity and reliability of the developed HPLC method. The compounds dehydrocostus lactone and costunolide were concentrated in the hexane fraction (222.08 and 65.07 µg/mg, respectively) and chloroform fraction (99.02 and 30.21 µg/mg, respectively), while the n-butanol fraction is a rich source of syringin (37.91 µg/mg) and 5-hydroxymethyl-2-furaldehyde (7.94 µg/mg). Further, the SRB assay was performed for the evaluation of anticancer activity using lung, colon, breast and prostate cancer cell lines. The hexane and chloroform fractions show excellent IC50 values of 3.37 ± 0.14 and 7.527 ± 0.18 µg/mL, respectively, against the prostate cancer cell line (PC-3).

Bioassay-Guided Isolation of Dehydrocostus Lactone from <em>Echinops kebericho</em> as a Leishmanicidal Drug

“…The 1 H NMR (500 MHz, CDCl3) showed a spectra of δ 1 H: 6.19 (1H, d, J = 3.5 Hz, H-13a), 5.46 (1H, d, J = 3.2 Hz, H-13b), 5.24 (1H, brs, J = 2.4, 1.5 Hz, H-15a), 5.04 (1H, brs, H-15b), 4.87 (1H, brs, H-14a), 4.79 (1H, brs, H-14b), 3.94 (1H, t, J = 9.3 Hz, H-6), 2.93 (1H , m, H-5), 2.89 -2.79 (2H, m, H-4, 7), 2.59 -2.41 (3H, m, H-3a, 9a, 3b), 2.22 (1H, m, H-9b ), 2.13 (1H, m, H-2a), 1.97 -1.79 (2H, m, H-2b,8a), 1.39 (1H, m, H-8b) (Figure 3). From the spectral analysis and comparisons with the literature [22] [23], the structure of Ek-cpd1 was characterized as dehydrocostus lactone, which is a sesquiterpene lactone with guaianolides class (Figure 4).…”