Background: The purpose of these studies was to determine the safety of a botanical treatment for supporting healthy liver function developed in Peru. The formulation, A4+, contains extracts of Curcuma longa L. rhizome (A4R), Cordia lutea Lam. flower (A4F) and Annona muricata L. leaf (A4L). The tests were used to support an application for a non-traditional Natural Health Product Licence from the Natural Health Product Directorate of Health Canada and future clinical trials.
Methods: Besides reviewing the scientific and clinical information from Peru on the ingredients and conducting an initial Ames test for mutagenicity, we analysed A4+ for its chemical profile and tested genotoxicity (micronucleus test) and general toxicity (28-day repeated dose).
Results: A4+ and extracts from the three plants provided distinctive chemical fingerprints. A4L contained acetogenins, requiring a second chromatographic method to produce a specific fingerprint. The Ames test proved positive at the highest concentration (5,000 μg/mL) but A4+ showed no evidence of genotoxicity in the more specific mouse micronucleus test. The 28-day repeated dose (general toxicity) study in rats showed no toxicity at 2,000 mg/kg.
Conclusions: We conclude that under the conditions of these studies, A4+ shows no evidence of toxicity at the levels indicated. A no observed adverse effect level (NOAEL) of 2,000 mg/kg was assigned.
Author contributions: HS directed the toxicology studies and drafted the manuscript. DS conducted the analytical chemistry. FG coordinated the study and manuscript preparation. SA provided medical and herbal expertise. AC assisted with background research and coordination of the project. JC invented the A4+ formulation and provided the ingredients for testing. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: We wish to acknowledge Sabell Corporation for financing these studies.
Employment or leadership: Drs José Cabanillas, Francis Green and Steven Aung are shareholders in Sabell Corp and members of Sabell’s scientific advisory committee. Drs. Hugh Semple and Duff Sloley are members of Sabell’s scientific advisory committee and contributed to the safety and efficacy PLA submission to Health Canada.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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