Effect of Altitude and Solvent on Psidium guajava Linn. leaves extracts: Phytochemical Analysis, Antioxidant, Cytotoxicity and Antimicrobial activity against Food Spoilage Microbes

Abstract

Background: Guava (Psidium guajava Linn.) has been traditionally used in the treatment of a wide range of diseases due to its rich content of secondary metabolites. This study was aimed to evaluate the effect of altitude and solvent systems on guava leaves crude extract’s phenolics and flavonoid content, antioxidant, antimicrobial, and its toxicity nature.Methods: Guava leaves were collected from three different geographical locations of Nepal while solvents with an increasing polarity index were used for extraction. The yield percentage of extracts was calculated. Total Phenolic Content, Total Flavonoid Content, and antioxidant activity were determined by the Folin-Ciocalteu method, Aluminium chloride colorimetric method, and DPPH (2,2’-Diphenyl-1-picrylhydrazyl) assay respectively. The quantification of fisetin and quercetin was performed using the HPLC with method validation. The antimicrobial activity of the extracts was tested against bacteria and fungus isolated from spoiled fruits and vegetables and identified through 16s and 18s rRNA sequencing. Finally, Brine Shrimp Lethality Assay (BSLA) was used for testing the toxicity of the extracts.Results: The phenolic and total flavonoid content was found to be higher in ethanol extract (331.84 mg GAE/g) and methanol extract (95.53 mg QE/g) from Kuleshwor respectively. Water extract of guava leaves from Kuleshwor (WGK) did not show significantly different antioxidant activity when compared to methanol and ethanol extracts. Fisetin and quercetin were higher in WGK (1.176 mg/100 g) and (10.967 mg/100 g) dry extract weight respectively. Antibacterial activity against food spoilage bacteria was dose dependent and found to be highest for all the extracts from different solvents and altitude at higher concentrations (80 mg/ml). Similarly, methanol and ethanol guava extracts from all locations showed antifungal activity against Geotrichum candidum RIBB-SCM43 and Geotrichum candidum RIBB-SCM44. WGK was found to be non-toxic.Conclusion: Our study concludes that the antioxidant and antimicrobial activity of WGK was found to be simlar with that of methanol and ethanol extracts of Bishnupur Katti and Mahajidiya. These results suggest the possibility of using water as a sustainable solvent to extract natural antioxidant and antimicrobial compounds which can further be used as natural preservatives to extend the shelf life of fruits and vegetables.