Evaluation of in vitro toxicity of common phytochemicals included in weight loss supplements using 1H NMR Spectroscopy

Emily C Davies; Garth Maker; Ian Musgrave; Sam Lodge

doi:10.1002/2211-5463.70170

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Evaluation of in vitro toxicity of common phytochemicals included in weight loss supplements using 1H NMR Spectroscopy

Journal article

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Evaluation of in vitro toxicity of common phytochemicals included in weight loss supplements using 1H NMR Spectroscopy

Emily C Davies, Garth Maker, Ian Musgrave and Sam Lodge

FEBS open bio, Early View

2025

DOI: https://doi.org/10.1002/2211-5463.70170

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Abstract

1 H NMR spectroscopy

Caco-2 cells

epigallocatechin-3,0-gallate

HepG2 cells

herbal weight loss supplements

oxidative stress Caco-2, colon carcinoma

CYP3A, cytochrome P450 3A

CYP450, cytochrome P450

D 2 O, Deuterium oxide

DILI, drug-induced liver injury

DMEM, Dulbecco's Modified Eagle Medium

DMSO, Dimethyl sulfoxide

EDTA, Ethylenediaminetetraacetic acid

EGCG, Epigallocatechin-3,0-gallate

FBS, fetal bovine serum

fc, fold change

FDA, Food and Drug Administration

GTE, green tea extract

HCA, hydroxycitric acid

HDS, herbal and dietary supplements

HepG2, human liver carcinoma

HFP, human Foods Program

HILI, herb-induced liver injury

MeOH, Methanol

MTT, Thiazolyl blue tetrazolium bromide

NMR, Nuclear Magnetic Resonance

OPLS-DA, orthogonal projection to latent structures discriminant analysis

PBS, Phosphate buffered saline

PCA, Principal component analysis

PQN, probabilistic quotient normalisation

R2X, model interpretation rate

ROS, reactive oxygen species

SEM, standard error of the mean

SSRIs, selective serotonin reuptake inhibitors

TCA, Tricarboxylic acid

TGA, Therapeutic Goods Administration

WHO, World Health Organisation

WLS, weight loss supplements

Herbal and dietary supplements (HDS) are popular among consumers seeking a 'natural' approach for improving their health; however, at present , there is a lack of evidence to support the claims of efficacy and safety for most of these products. Herbal weight loss supplements (WLS) are a group of HDS that are frequently implicated in cases of toxicity; however, the causative substances often remain unknown due to the complex chemical nature of such supplements. This study aimed to analyse the in vitro safety (in human liver carcinoma (HepG2) cells and colon carcinoma (Caco-2) cells) of 12 active compounds commonly found in WLS, first with safety screening using the MTT cytotoxicity assay, followed by metabolic profiling with 1 H NMR spectroscopy. Of the phytochemicals evaluated, epigallocatechin-3,0-gallate (EGCG) was the only compound that caused a significant reduction in the viability of both cell lines (25.3% in HepG2 cells and 18.5% in Caco-2 cells), and this decrease was potentiated by CYP450 induction with rifampicin. Subsequent 1 H NMR analysis showed changes in key metabolites such as amines, amino acids, carboxylic acids, and glucose that were indicative of protein degradation and disrupted energy and lipid metabolism. While the remaining 11 active compounds analysed did not demonstrate significant toxicity in isolation, these require further assessment to determine their safety when used in combination with other phytochemicals. Given that the majority of WLS contain multiple herbal ingredients, each with a complex chemical composition, it is important to understand the role of interactions in adverse events. Abbreviations ANOVA, analysis of variance; ARTG, Australian Register of Therapeutic Goods; AUROC, area under the receiver operator curve; BBI, broadband decoupling inverse;

Details

Title: Evaluation of in vitro toxicity of common phytochemicals included in weight loss supplements using 1H NMR Spectroscopy
Authors/Creators: Emily C Davies - Murdoch University
Garth Maker - Murdoch University, Centre for Computational and Systems Medicine
Ian Musgrave - The University of Adelaide
Sam Lodge (Author) - Murdoch University, Centre for Computational and Systems Medicine
Publication Details: FEBS open bio, Early View
Publisher: John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.
Number of pages: 15
Identifiers: 991005838266607891
Murdoch Affiliation: School of Medical, Molecular and Forensic Sciences; Centre for Computational and Systems Medicine
Language: English
Resource Type: Journal article

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