Comprehensive HPTLC Fingerprinting and Quality Evaluation of Punicalagin in Pomegranate (Punica granatum L.) Aril Juice

Objectives:

Pomegranate (Punica granatum) is a well-known source of bioactive phytochemicals, particularly punicalagin, which contributes to its antioxidant, anti-inflammatory, and antimicrobial properties.

Materials and Methods:

In this study, a robust High-Performance Thin-Layer Chromatography (HPTLC) method was developed to fingerprint and quantify punicalagin in aril juice.

Results:

The optimized mobile phase-chloroform: ethyl acetate: formic acid: methanol (4.0:5.2:0.6:0.2, v/v/v/v)-produced sharp, well-resolved bands, and densitometric scanning at 620 nm revealed a distinct punicalagin peak at (Retention factor) R_f =0.58±0.03, accounting for 5.32% of the total chromatographic area. Microbial evaluation confirmed the absence of E. coli, S. aureus, Salmonella, Candida albicans, and Shigella, with total aerobic and yeast/mould counts below 10 (Colony Forming Units) CFU/g. Elemental screening revealed trace levels of Pb, Cd, and Hg well below pharmacopeial limits, and nutritionally relevant minerals such as iron, zinc, and copper were present in appreciable amounts.

Conclusion:

These findings establish a reproducible HPTLC fingerprint for pomegranate aril juice and demonstrate that integrated chemical, microbial, and elemental profiling provides a comprehensive approach for quality control and authentication of pomegranate-derived nutraceuticals.