Phytochemical evaluation and preliminary formulation development of Acalypha indica (Kuppameniya) extract as a cat mood enhancer

Abstract

In Sri Lanka, Acalypha indica known to influence the behaviour of domestic cats, often inducing playful and euphoric responses. This study aimed to investigate the plant at the laboratory scale by identifying its bioactive constituents with potential cat-attractant properties and developing a preliminary formulation as a cat mood enhancer. Fresh roots and leaves of the plant were extracted separately with methanol. Phytochemical constituents present in methanolic extracts were examined by standard chemical tests and UV spectroscopic analysis. The possible cat-attracting compounds in the root extract were determined using Gas Chromatography-Mass Spectrometry (GC-MS). The study also evaluated the safety of the crude extract through zebrafish embryo toxicity assay for the identification of LC50 value. Based on the toxicity findings, preliminary formulations of different strengths were developed. The formulation used polyethylene glycol as a fixative, with glycerin and ethanol as preservatives to stabilise the volatile cat-attractant compounds. The highest non-toxic concentration formulation was evaluated by short-term stability study through Solid-Phase Microextraction (SPME) and physical testing. Phytochemical screening revealed the presence of alkaloids, phenols, flavonoids, terpenoids, tannins, and steroids. GC-MS analysis revealed the presence of two of the known cat attractants, Iridomyrmecin and Dihydronepetalactone, in the root extract. LC50 of the crude extract was determined to be 10.71% w/w. SPME analysis after one month revealed a loss of 50.95% in Iridomyrmecin and 58.84% in Dihydronepetalactone levels, indicating the need for formulation optimization. Furthermore, direct animal trials will be essential to verify efficacy and safety prior to any commercial application. In conclusion, the identification of well-known cat-attracting compounds Iridomyrmecin and Dihydronepetalactone highlights the potential of Acalypha indica for the regulation of feline behaviour as per traditional practice. This study focused solely on the preliminary formulation and laboratory evaluation of Acalypha indica using methanol for extraction and phytochemical characterisation. As the formulation has not yet been tested in animals, methanol was used in this preliminary study. However, for future animal-targeted applications, only safe and non-toxic polar solvents should be employed.