Characterization and anticancer potential of Withania somnifera fruit bioactives (a native species to Pakistan) using gas chromatography-mass spectrometer, nuclear magnetic resonance and liquid chromatography-mass spectrometry-electrospray ionization

doi:10.19852/j.cnki.jtcm.2022.06.006

Abstract

Abstract:

INTRODUCTION: Withania somnifera (W. somnifera) is a plant with remarkable pharmacological properties. The plant has an impressive profile of medicinal uses in the folk medicine system of several civilizations.

AIM: This comprehensive study is aimed to characterize phytochemicals in fruit of W. somnifera and tested for in vitro anticancer potential to find out active candidate in disease prevention and treatment.

METHODS: The bioactive components from W. somn-ifera fruit were extracted with polar and non-polar solvents. Anticancer potential of the isolated bioactive was assessed against different cancer cell lines through MTT assay and Incucytes imaging analysis. The extracts were characterized for secondary metabolites using GC-MS (gas chromatography-mass spectrometer), LCMS (liquid chromatography-mass spectrometry)-ESI (electrospray Ionization) and 1H-NMR (electrospray Ionization) techniques.

RESULTS: Both freeze-dried and rotary evaporator con-densed extracts exhibited anticancer potential against MDA-MB-231, MCF7- SKOV3 and SKBR3 cell lines. The tested extracts have cell growth inhibition potential ag-ainst mammalian cancer cell line. Hexacosanedioic acid purified from n-hexane extract through HPLC was inves-tigated for its cytotoxicity against breast cancer cell line SKBR3 by using Incucytes imaging analysis. Conclusion: We found that a variety of bioactive compounds existed in this plant. One identified compound that was not investigated for cytotoxicity in previous studies was purified and its application showed cytotoxicity on breast cancer cell lines. A number of bioactive identified from W. somnifera fruit may have an effective potential for development into chemotherapy drugs.

Key words: phytochemicals, plants, medicinal, anti-cancer activities, purification, state-of-the-art techniques

Qudsia Tabassam, Tahir Mehmood, Sibtain Ahmed, Farooq Anwar, Abdul Rauf Raza. Characterization and anticancer potential of Withania somnifera fruit bioactives (a native species to Pakistan) using gas chromatography-mass spectrometer, nuclear magnetic resonance and liquid chromatography-mass spectrometry-electrospray ionization[J]. Journal of Traditional Chinese Medicine, 2022, 42(6): 908-918.

Figures/Tables 5

Figure 1 Cell viability of four cancer cell lines against treated extracts, positive control drugs and negative control media with corresponding 1% DMSO concentration. Values are reported as mean ± SD (n = 3) of three separate experiments. A: SKBR3; B: MDA-MB-231; C: MCF7; D: SKOV3.

Figure 2 Ten compounds

Figure 3 Anticancer activity of Hexacosanedioic acid towards breast cancer cell line

Figure 4 cell experiment result A: % Confluency after 3 days treatment; B: negative control; C: indicating with drug 25 μg/mL conc; Sample highest concentration 50 μg/mL.

Table 3 Hexacosanedioic acid mass and various confirmatory parameters

Actual m/z (amu)	Ion formula	Score	Compound	Calculated m/z (amu)	Mean mass Err (mmu)	Mean error (ppm)	M Sigma	Relative double bond		eConf	N-Rule	XR Score
427.37864	C₂₆H₅₁O₄	100	Hexacosane-dioic acid	427.3782	-4	-1.4	89.1		2	even	ok	n.a

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