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International Journal of Pharmaceutical InvestigationNOW IN
Article

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

1, 1★, 2, 2, 1★ Corresponding author

  1. 1Department of Pharmacology, Saveetha Institute of Medical and Technical Sciences, Chennai, Tamil Nadu, INDIA.
  2. 2Department of Pharmacology, M V J Medical College and Research Hospital, Bengaluru, Karnataka, INDIA.

CORRESPONDENCE

Thirunavukkarasu Jayaraman

Pharmacology, Saveetha Institute of Medical and Technical Sciences, Chennai-602105, Tamil Nadu, INDIA.

thirunavukkarasu75@outlook.com

ORCID: 0009-0004-0333-4599

Received: 11-06-2026; Revised: 24-07-2026; Accepted: 07-09-2026.

Volume 17, Issue 1 · pp. 249–255 · 2027 · DOI: 10.5530/ijpi.20270024

View on IJPI original site ↗

ABSTRACT

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) Rf =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.

KEYWORDS

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FULL TEXT

INTRODUCTION

Medicinal plants are an indispensable source of bioactive compounds that contribute to human health by preventing and managing chronic diseases. These plant-derived phytochemicals-including phenolic compounds, flavonoids, tannins, and alkaloids-exert diverse biological effects such as antioxidant, anti-inflammatory, antimicrobial, and anticancer activities, making them vital candidates for drug discovery and development (Mo et al., 2022; Singh et al., 2018). Rising antimicrobial resistance has renewed global interest in exploring plant-based therapeutics as alternative or adjunct treatments, offering solutions where conventional antibiotics have become ineffective (DEBIB et al., 2022; Kiran et al., 2024; Maslov et al., 2025).

Pomegranate (Punica granatum L.) is a widely consumed fruit valued for its nutritional and therapeutic properties. Its peel, seeds, and arils are rich in phenolic compounds-including flavonoids (anthocyanins, anthoxanthins), tannins (ellagitannins such as punicalagin, punicalin, and pedunculagin), and phenolic acids (chlorogenic, caffeic, syringic, sinapic, p-coumaric, ferulic, ellagic, gallic, and cinnamic acids)-along with dietary fiber, minerals, and vitamins (Du et al., 2025; Ranjha et al., 2023; Siddiqui et al., 2024; Singh et al., 2018). Among these, punicalagin is the most abundant and pharmacologically significant ellagitannin, exhibiting potent antioxidant, anti-inflammatory, anticancer, cardioprotective, and antimicrobial activities (Shabir et al., 2024; Venusova et al., 2021; Wang et al., 2024; Xu et al., 2021). Mechanistic studies demonstrate that punicalagin modulates key signaling pathways-including Nuclear Factor kappa B (NF-Κb), Mitogen-Activated Protein Kinase (MAPK) and Phosphoinositide 3-kinase/Protein Kinase B/Mammalian Target of Rapamycin (PI3K/Akt/mTOR)-attenuates oxidative stress, induces apoptosis in malignant cells, and improves immune and endothelial function (Chen et al., 2023; Ramlagan et al., 2022; Tang et al., 2022). These multifaceted bioactivities make punicalagin a promising candidate for nutraceutical and pharmaceutical applications, though its clinical translation requires consistent product characterization and standardized dosing (Shabir et al., 2024; Xu et al., 2021).

Pomegranate juice, particularly from the arils, is a major dietary source of anthocyanins and other polyphenols, contributing to its strong antioxidant capacity (Habib et al., 2023; Karagecili et al., 2023). In vitro and in vivo studies have demonstrated its ability to inhibit pathogenic bacteria, including Multi Drug-Resistant (MDR) strains such as Methicillin-Resistant Staphylococcus aureus (MRSA) and Extended-Spectrum Β-Lactamase (ESBL)-producing Enterobacteriaceae, by disrupting microbial growth and resistance mechanisms (Alghamdi et al., 2024; Gosset-Erard et al., 2021; Maslov et al., 2025). These findings support the potential of pomegranate-derived products as natural antimicrobials, functional foods, and adjunct therapies to combat antibiotic resistance.

Reliable quantification and profiling of bioactive compounds are essential for ensuring the authenticity, reproducibility, and quality of pomegranate-based products. High-Performance Thin-Layer Chromatography (HPTLC) is a robust and cost-effective technique that allows simultaneous analysis of multiple phytochemicals with minimal sample preparation, producing a reproducible chemical “fingerprint” for qualitative and quantitative assessments (Venkatesham et al., 2021). HPTLC is particularly suitable for detecting marker compounds such as punicalagin, distinguishing genuine from adulterated products, and enabling high-throughput quality control in both research and industry (Farag et al., 2020; Riolo et al., 2025; Skrt et al., 2022). While High-Performance Liquid Chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry MDR (LC-MS0 provide higher resolution and sensitivity, HPTLC offers a rapid and economical alternative for routine screening, especially where large-scale analysis is required (Farag et al., 2020; Riolo et al., 2025; Skrt et al., 2022).

Despite growing evidence of pomegranate’s therapeutic potential, several gaps remain. Quantitative standardization of punicalagin and other bioactive compounds across different juice preparations is inconsistent due to variability in plant parts, extraction methods, and analytical protocols (Magangana et al., 2020; Yisimayili and Chao, 2022). The lack of validated, comprehensive fingerprinting methods hinders reliable product comparison and clinical translation (Aziz et al., 2020; Laurindo et al., 2022). Large-scale, well-controlled clinical trials are still needed to substantiate health claims (Mohan et al., 2024; Mokgalaboni et al., 2023).

Therefore, the present study aims to develop and validate a HPTLC-based fingerprinting method for the quantification of punicalagin in pomegranate aril juice. This standardized approach will facilitate routine quality control, ensure product authenticity, and enable reproducibility in research and nutraceutical applications.

MATERIALS AND METHODS

Plant Material and Sample Collection

Fresh pomegranate (Punica granatum L.) fruits were collected from mature trees. Only fruits without visible defects, decay, or microbial contamination were selected to ensure sample integrity. The fruits were washed twice with Double-Distilled Water (ddH2O) and disinfected using a 150 µL/L sodium hypochlorite solution for 1 min, followed by thorough rinsing with sterile ddH2O to remove residual chlorine (Cite: Standard disinfection and preparation protocols). The arils were manually separated from the peel and seeds under aseptic conditions and processed immediately for juice extraction.

Sample Extraction

Approximately 1 kg of fresh pomegranate arils was macerated with 1 L of a hydroalcoholic solvent mixture (50:50, v/v, distilled water: methanol) to obtain a homogeneous slurry. The mixture was filtered through a double-layered muslin cloth to remove seeds and coarse pulp. The filtrate was concentrated to dryness using a water bath at controlled temperature (<50°C) to prevent thermal degradation of phenolic compounds. The resulting dry extract was stored in a desiccator until constant weight was achieved and subsequently weighed to determine the yield.

Preparation of Test Solution

1 g of the dried pomegranate extract was dissolved in 10 mL of HPTLC-grade methanol to prepare a stock solution. Aliquots of 3, 6, and 9 μL from this stock solution were used for HPTLC analysis. This methanolic extract served as the test solution for fingerprinting and quantitative determination of punicalagin (Bhargava et al., 2021).

HPTLC Fingerprinting and Quantification

The schematic plan of HPTLC finger printing was depicted in Figure 1.

Figure 1: Schematic presentation of Workflow for the preparation, HPTLC fingerprinting, quantitative analysis of punicalagin, and microbial/elemental assessment of pomegranate (Punica granatum) aril juice.

Chromatographic Conditions

HPTLC analysis was achieved on pre-coated silica gel 60 F254 aluminum plates (Merck, Germany). Using a CAMAG Linomat 5 applicator, 3, 6, and 9 μL of the methanol extract were applied as 7 mm-wide bands with a constant application rate under a nitrogen stream. The chromatograms were developed in a twin-trough glass chamber pre-saturated with a mobile phase consisting of chloroform: ethyl acetate: formic acid: methanol in the ratio of 4.0:5.2:0.6:0.2 (v/v/v/v). After development, plates were air-dried and visualized under short-wave (Ultraviolet) UV (254 nm) and long-wave UV (366 nm). Plates were then derivatized with anisaldehyde-sulphuric acid reagent, heated at 105°C until color development, and scanned densitometrically at 620 nm using a CAMAG Scanner 4 with winCATS software.

Statistical Analysis

Quantitative data obtained from densitometric scanning were expressed as mean ± Standard Deviation (SD). All HPTLC analyses were performed in triplicate to ensure reproducibility. Retention factor (Rf) values and peak areas were calculated using winCATS software (CAMAG, Switzerland).

Microbial and Elemental Analysis

Microbial analysis of pomegranate juice was conducted according to pharmacopeial guidelines (Cite: Indian/United states/British Pharmacopoeia). Tests included Total Aerobic Microbial Count (TAMC), Total Yeast and Mold Count (TYMC), and presence/absence of common foodborne pathogens (E. coli, Salmonella spp., S. aureus). Elemental examination was performed using standard pharmacopeial procedures to regulate the existence and concentration of essential minerals, including potassium, sodium, calcium, and trace elements (Popovici et al., 2021).

RESULTS

HPTLC Fingerprinting Profile

The methanolic extract of Punica granatum aril juice was successfully analyzed using High-Performance Thin-Layer Chromatography (HPTLC) to generate a characteristic fingerprint profile. The optimized mobile phase system-chloroform: ethyl acetate: formic acid: methanol (4.0:5.2:0.6:0.2, v/v/v/v)-produced well-resolved, sharp, and symmetrical peaks without overlapping bands. The densitometric scan at 620 nm revealed four distinct peaks, with Peak 3 corresponding to punicalagin (Rf=0.58±0.03).

This peak accounted for 5.32% of the total chromatographic area, confirming its significant presence in the extract (Figure 2 and Table 1).

Figure 2: HPTLC densitometric chromatogram of Punica granatum aril juice extract (6 µL) scanned at 620 nm, showing four major peaks. Peak 3 (Rf=0.58±0.03) corresponds to punicalagin.
Table 1: HPTLC Densitometric Data of Punica granatum hydroalcoholic Extract (6 µL).
Peak No.Start Position (Rf)Max Position (Rf)End Position (Rf)Max Height (AU)Area (AU)Area %
10.010.050.08422.06331.178.16
20.110.160.1938.6734.49.07
3 (Punicalagin)0.490.550.5914.9431.25.32
40.690.720.7814.3603.47.45

Chromatogram and Photo-Documentation

The HPTLC chromatogram displayed clear separation of components, as indicated by discrete peaks across the Rf range (0.01–0.72). Photodocumentation of the developed plates under short-wave UV (254 nm), long-wave UV (366 nm), and white light (after derivatization with anisaldehyde-sulphuric acid) confirmed the presence of multiple phytoconstituents with distinct fluorescence and color intensities (Figure 3). The band corresponding to punicalagin was prominent across all applied concentrations (3, 6, and 9 μL), supporting the specificity and reproducibility of the method.

Figure 3: HPTLC photodocumentation of Punica granatum hydroalcoholic extract under (A) short-wave UV (254 nm), (B) long-wave UV (366 nm), and (C) white light after derivatization with anisaldehyde-sulphuric acid. Tracks: 1=3 µL, 2=6 µL, 3=9 µL.

Specificity and Quantification

Specificity of the HPTLC method was verified by comparing the Rf value and densitometric profile of the sample with the punicalagin reference standard under identical chromatographic conditions. The results demonstrated no co-elution with other constituents, confirming the method’s selectivity for punicalagin quantification.

Densitometric data analysis

The four distinct peaks measured, with Peak 1 accounting up most (78.16% area). Punicalagin occupied 5.32% of the total peak area and was found with an Rf value of 0.55 in Peak 3.

Densitometric Profiling extract

After development chromatographic development, plates were derivatized and scanned densitometrically at 620 nm to assess the (Start, Maximum End) Rf values, maximum peak height and relative percentage of identified compounds. The four well defined peaks were found showing Rf values of 0.05, 0.16, 0.55 and 0.72 indicates the confirming different phytochemical components in the prepared extract (Figure 4).

Figure 4: HPTLC densitometric peak table of the hydroalcoholic extract of Punica granatum fruit arils at 620 nm.

Microbial Quality Assessment

Microbiological analysis of the dried pomegranate aril juice extract was performed according to pharmacopeial guidelines. The results specified that the sample met the microbiological quality criteria for herbal preparations. Total Aerobic Bacterial Count (TAMC) and Total Yeast and Mold Count (TYMC) were both found to be below 10 CFU/g, indicating minimal microbial load. No pathogenic organisms-including Escherichia coli, Staphylococcus aureus, Salmonella spp., Candida albicans, or Shigella spp.-were detected in the sample, confirming its microbiological safety for consumption (Table 2).

Table 2: Microbial Quality Analysis of Punica granatum Aril Juice Extract.
Sl. No.Test ParameterObserved Value
1.Total Aerobic Count (CFU/g)<10
2.Yeast and Mould Count (CFU/g)<10
3.Escherichia coli (CFU/g)Negative
4.Staphylococcus aureus (CFU/g)Negative
5.Salmonella spp. (CFU/g)Negative
6.Candida albicans (CFU/g)Negative
7.Shigella spp. (CFU/g)Negative

Elemental Analysis

Elemental screening of the dried pomegranate aril juice powder was performed according to pharmacopeial guidelines to determine the presence of essential minerals and potential toxic heavy metals. The results demonstrated that the concentrations of lead, cadmium, and mercury were well below permissible limits, indicating no risk of heavy metal contamination. Among the essential elements, iron and zinc were found in appreciable amounts, followed by copper, suggesting that the juice contributes beneficial micronutrients (Table 3).

Table 3: Elemental Analysis of Punica granatum Aril Juice Powder.
Sl. No.ElementObserved Value (ppm)
1.Lead (Pb)0.23
2.Cadmium (Cd)0.004
3.Mercury (Hg)0.002
4.Copper (Cu)9.077
5.Iron (Fe)39.8
6.Zinc (Zn)12.34

DISCUSSION

Chromatographic fingerprinting provides a unique chemical profile of complex matrices and is widely recognized as a powerful tool for identity testing and quality assessment of herbal medicines and food products (Meena et al., 2021). In the present study, an optimized High-Performance Thin-Layer Chromatography (HPTLC) method was developed for the fingerprinting and quantification of punicalagin in Punica granatum aril juice. The chosen mobile phase, chloroform: ethyl acetate: formic acid: methanol (4.0:5.2:0.6:0.2, v/v/v/v), produced shawell-resolved bands without interference from other phytoconstituents, confirming the specificity of the system. Densitometric analysis revealed four major peaks, with the peak corresponding to punicalagin appearing at an Rf value of 0.58±0.03, accounting for 5.32% of the total chromatographic area. Reporting the Rf value is particularly significant because most published HPTLC studies on punicalagin quantification focus on concentration data but omit Rf information, which is essential for reproducibility and inter-laboratory comparisons (Shahkoomahally et al., 2023). The presence of a distinct punicalagin band across all applied concentrations (3, 6, and 9 μL) further supports the robustness of the established method and its suitability for repetitive quality control applications.

Previous reports have quantified punicalagin in pomegranate peel and commercial herbal preparations using HPTLC, with concentrations ranging from 807.6 μg to 3.207 mg/g of sample (Gadkari and Daharwal, 2022). whereas most recent studies employ HPLC or UHPLC methods, reporting 5.2–130.32 μg/mL in peel extracts and up to 238.7 μg/mL in juice, depending on the cultivar and extraction method (Kumar et al., 2023; Maharana et al., 2022; Mahmood and Jabar, 2023). Although direct comparison of area percentages with concentration data is challenging due to methodological differences, the present study demonstrates that HPTLC can reliably detect and quantify punicalagin in aril juice, offering a cost-effective screening tool prior to more advanced HPLC or LC-MS-based analysis. This is consistent with WHO and pharmacopeial recommendations that advocate the use of HPTLC as a reproducible and regulatory-compliant tool for authentication, standardization, and detection of adulteration in herbal materials (El-Waey et al., 2023; Frommenwiler et al., 2022).

Punicalagin is one of the most pharmacologically significant ellagitannins in pomegranate, exhibiting antioxidant, anti-inflammatory, anticancer, cardioprotective, and antimicrobial properties (10,11,12). It modulates several key signaling pathways, including NF-κB, MAPK, and PI3K/Akt/mTOR, reduces oxidative stress, and improves immune and endothelial function (Chen et al., 2023; Ramlagan et al., 2022; Tang et al., 2022). Therefore, its detection and quantification are critical for establishing the nutraceutical value of pomegranate juice and ensuring batch-to-batch consistency in commercial formulations. The results of microbial analysis further strengthen the quality assessment of the juice powder, as total aerobic count and yeast/mold count were <10 CFU/g and no pathogens (Escherichia coli, Staphylococcus aureus, Salmonella spp., Candida albicans, or Shigella spp.) were detected, these findings indicate great microbial quality and are in accordance with better microbial resilience commonly associated with freeze-dried fruit powders with low moisture levels (Howell and D’Souza, 2013). Similarly, elemental analysis revealed that concentrations of toxic heavy metals (Pb, Cd, Hg) were well below pharmacopeial limits, while nutritionally relevant minerals such as iron, zinc, and copper were present in appreciable amounts. These findings are in line with reports that identify potassium and iron as dominant minerals in pomegranate juice and confirm the nutritional relevance of the analyzed material (Nuncio-Jáuregui et al., 2015).

Collectively, these results demonstrate that HPTLC fingerprinting coupled with microbial and elemental profiling provides a comprehensive approach for the authentication and quality control of pomegranate aril juice. The reported Rf value for punicalagin can serve as a reference parameter for future studies, and the established fingerprint profile offers a baseline for comparison with other varieties or commercial products. Future work should include full validation of the method-covering precision, accuracy, Limit of Detection (LOD), and Limit of Quantification (LOQ)-and comparative studies using HPLC or LC-MS to benchmark the sensitivity and specificity of the approach. Large-scale screening of pomegranate juices from different cultivars and processing methods would further strengthen the applicability of this technique for routine quality assurance and support the development of standardized, evidence-based nutraceutical formulations.

CONCLUSION

This investigation successfully developed an HPTLC-based fingerprinting method for the qualitative and quantitative analysis of punicalagin in Punica granatum aril juice, reporting an Rf value of 0.58±0.03 and confirming its significant presence through densitometric analysis. The combined use of microbial and elemental profiling further established the safety and nutritional relevance of the sample, providing a comprehensive quality assessment framework. These findings support the use of HPTLC as a robust, cost-effective tool for routine authentication, standardization, and quality control of pomegranate-derived nutraceuticals. Future work focusing on full analytical validation and comparative studies with HPLC/LC-MS will enhance the reliability and regulatory acceptance of this approach.

ACKNOWLEDGEMENT

We thank the Department of pharmaceutical chemistry and Pharmacognosy SDM Centre for Research in Ayurveda and Allied Sciences, Udupi, Karnataka, Saveetha Institute of Medical and Technical Sciences and MVJ Medical College and Research Hospital Hoskote Karnataka. For providing necessary laboratory facilities, advices and technical support.

ABBREVETIONS

CFU
Colony-Forming Unit
ddH2O
Double-distilled water
ESBL
Extended-Spectrum β-Lactamase
HPLC
High-Performance Liquid Chromatography
HPTLC
High-Performance Thin-Layer Chromatography
LC-MS
Liquid Chromatography-Mass Spectrometry
LOD
Limit of Detection
LOQ
Limit of Quantification
MAPK
Mitogen-Activated Protein Kinase
MDR
Multidrug-Resistant
MRSA
Methicillin-Resistant Staphylococcus aureus
NF-κB
Nuclear Factor kappa B
PI3K/Akt/mTOR
Phosphoinositide 3-kinase/Protein Kinase B/Mammalian Target of Rapamycin
Rf
Retention factor
SD
Standard Deviation
TAMC
Total Aerobic Microbial Count
TYMC
Total Yeast and Mold Count
USP/BP
United States Pharmacopeia/British Pharmacopoeia
UV
Ultraviolet.

CONFLICT OF INTEREST

The authors declare that there is no conflict of interest.

FUNDING

This work is not resourced by any private or public sectors.

AUTHORS CONTRIBUTION

RPC: Conceptualization, study designing, data collection, manuscript drafting and resources. BU: Conceptualization and study design. AKAK: Manuscript drafting and data analysis. TJ: Supervision and approval of final draft of manuscript.

AVAILABILITY OF DATA AND MATERIAL

All data has been depicted in the study and for any further clarifications, the required can be obtained from corresponding author upon request.

ETHICAL APPROVAL AND CONSENT TO PARTICIPATE

This study was done in an in vitro laboratory environment not affecting any person or animal in any manner.

CLINICAL TRIAL REGISTRATION

Not applicable

REFERENCES

As published

Showing references and in-text citations exactly as published.

  1. 1.Alghamdi, M. A.; Al-Sarraj, F.; Alamshani, W. H. et al. Antibacterial power of Pomegranate extracts against Beta-Lactamase producing Escherichia coli. Caryologia. 2024;77(1):83–99. https://doi.org/10.36253/caryologia-2356CrossrefGOOGLE SCHOLAR
  2. 2.Aziz, Z.; Huin, W. K.; Hisham, M. D. B.; Ng, J. X.. Effects of pomegranate on lipid profiles: A systematic review of randomised controlled trials. Complementary Therapies in Medicine. 2020;48. https://doi.org/10.1016/j.ctim.2019.102236CrossrefGOOGLE SCHOLAR
  3. 3.Bhargava, A.; Shrivastava, P.; Tilwari, A.. HPTLC analysis of Fumaria parviflora (Lam.) methanolic extract of whole plant. Future Journal of Pharmaceutical Sciences. 2021;7(1):1. https://doi.org/10.1186/s43094-020-00150-xCrossrefGOOGLE SCHOLAR
  4. 4.Chen, P.; Chen, F.; Lei, J.; Zhou, B.. Pomegranate polyphenol punicalagin improves learning memory deficits, redox homeostasis, and neuroinflammation in aging mice. Phytotherapy Research. 2023;37(9):3655–3674. https://doi.org/10.1002/ptr.7848CrossrefGOOGLE SCHOLAR
  5. 5.DEBIB, A.; Menadi, S.; Sahnouni, F.; Boukhatem, M. N.; Ouahchia, C.; Kaced, A.. Bacterial Inhibitory Effect Of Algerian Pomegranate (Punica granatum L.) Extracts (Peel, Juice, And Seed) Against Multidrug Resistant Bacteria. Journal of Microbiology, Biotechnology and Food Sciences. 2022;11(4). https://doi.org/10.55251/jmbfs.4622CrossrefGOOGLE SCHOLAR
  6. 6.Du, J.; Wang, H.; Zhong, L.; Wei, S.; Min, X. et al. Bioactivity and biomedical applications of pomegranate peel extract: a comprehensive revie. Frontiers in Pharmacology. 2025;16. https://doi.org/10.3389/fphar.2025.1569141CrossrefGOOGLE SCHOLAR
  7. 7.El-Waey, A. A.; Abdel-Salam, R. A.; Hadad, G. M.; El-Gindy, A.. Eco friendly stability indicating HPTLC method for simultaneous determination of sofosbuvir and ledipasvir in pharmaceutical tablets and HPTLC-MS characterization of their degradation products. Microchemical Journal. 2023;186(6). https://doi.org/10.1016/j.microc.2022.108324CrossrefGOOGLE SCHOLAR
  8. 8.Farag, R. S.; Abdel-Latif, M. S.; Baky, H. H. A.; El; Tawfeek, L. S.. Phytochemical screening and antioxidant activity of some medicinal plants’ crude juices. Biotechnology Reports. 2020;28. https://doi.org/10.1016/j.btre.2020.e00536CrossrefGOOGLE SCHOLAR
  9. 9.Frommenwiler, D. A.; Reich, E.; Sharaf, M. H. M.; Cañigueral, S.; Etheridge, C. J.. Investigation of market herbal products regulated under different categories: How can HPTLC help to detect quality problems?. Frontiers in Pharmacology. 2022;13. https://doi.org/10.3389/fphar.2022.925298CrossrefGOOGLE SCHOLAR
  10. 10.Gadkari, P.; Daharwal, S. J.. Quantification of Punicalagin in Pomegranate Peels from High-performance Thin-layer Chromatography. Biomedical and Biotechnology Research Journal (BBRJ). 2022;6(4):586–590. https://doi.org/10.4103/bbrj.bbrj_312_22CrossrefGOOGLE SCHOLAR
  11. 11.Gosset-Erard, C.; Zhao, M.; Lordel-Madeleine, S.; Ennahar, S.. Identification of punicalagin as the bioactive compound behind the antimicrobial activity of pomegranate (Punica granatum L.) peels. Food Chemistry. 2021;352(9):129396. https://doi.org/10.1016/j.foodchem.2021.129396CrossrefGOOGLE SCHOLAR
  12. 12.Habib, H. M.; El-Gendi, H.; El-Fakharany, E. M.; El-Ziney, M. G. et al. Antioxidant, Anti-Inflammatory, Antimicrobial, and Anticancer Activities of Pomegranate Juice Concentrate. Nutrients. 2023;15(12):2709. https://doi.org/10.3390/nu15122709CrossrefGOOGLE SCHOLAR
  13. 13.Howell, A. B.; D’Souza, D. H.. The Pomegranate: Effects on Bacteria and Viruses That Influence Human Health. Evidence-Based Complementary and Alternative Medicine. 2013;2013(1):1–11. https://doi.org/10.1155/2013/606212CrossrefGOOGLE SCHOLAR
  14. 14.Karagecili, H.; İzol, E.; Kirecci, E.; Gulcin, İ.. Determination of Antioxidant, Anti-Alzheimer, Antidiabetic, Antiglaucoma and Antimicrobial Effects of Zivzik Pomegranate (Punica granatum)-A Chemical Profiling by LC-MS/MS. Life. 2023;13(3). https://doi.org/10.3390/life13030735CrossrefGOOGLE SCHOLAR
  15. 15.Kiran, S.; Tariq, A.; Iqbal, S.; Naseem, Z.; Siddique, W. et al. Punicalagin, a pomegranate polyphenol sensitizes the activity of antibiotics against three MDR pathogens of the Enterobacteriaceae. BMC Complementary Medicine and Therapies. 2024;24(1):93. https://doi.org/10.1186/s12906-024-04376-7CrossrefGOOGLE SCHOLAR
  16. 16.Kumar, A.; P, N.; Kumar, M.; Jose, A.; Tomer, V.; Oz, E. et al. Major Phytochemicals: Recent Advances in Health Benefits and Extraction Method. Molecules. 2023;28(2):887. https://doi.org/10.3390/molecules28020887CrossrefGOOGLE SCHOLAR
  17. 17.Laurindo, L. F.; Barbalho, S. M.; Marquess, A. R.; de Souza Grecco, A. I.; de Alvares Goulart, R. et al. Pomegranate (Punica granatum L.) and Metabolic Syndrome Risk Factors and Outcomes: A Systematic Review of Clinical Studies. Nutrients. 2022;14(8):1665. https://doi.org/10.3390/nu14081665CrossrefGOOGLE SCHOLAR
  18. 18.Magangana, T. P.; Makunga, N. P.; Fawole, O. A.; Opara, U. L.. Processing Factors Affecting the Phytochemical and Nutritional Properties of Pomegranate (Punica granatum L.) Peel Waste: A Review. Molecules. 2020;25(20):4690. https://doi.org/10.3390/molecules25204690CrossrefGOOGLE SCHOLAR
  19. 19.Maharana, S.; Mangaonkar, K.; Phanse, M.. Microscopic and HPTLC Fingerprint Analysis a Tool for Authentication and Quality Control of Nelumbo nucifera. Chemistry Africa. 2022;5(3):663–672. https://doi.org/10.1007/s42250-022-00338-zCrossrefGOOGLE SCHOLAR
  20. 20.Mahmood, A.; Jabar, H.. Characterization of biochemical compounds in different accessions of pomegranate (Punica granatum L.) peels in Iraq. Passer Journal of Basic and Applied Sciences. 2023;5(2):382–390. https://doi.org/10.24271/psr.2023.409738.1363CrossrefGOOGLE SCHOLAR
  21. 21.Maslov, O.; Komisarenko, M.; Ponomarenko, S.; Osolodchenko, T.; Kolisnyk, S.. Analysis of Composition and Antimicrobial Activity Of Pomegranate (Punica Granatum) Fruit Extract Against Gram-Negative Multi-Drug Resistant Bacteria from Clinical Isolates. Journal of Microbiology, Biotechnology and Food Sciences. 2025;14(6). https://doi.org/10.55251/jmbfs.11389CrossrefGOOGLE SCHOLAR
  22. 22.Meena, A. K.; Rekha, P.; Perumal, A.; Ilavarasan, R.; Singh, R.; Srikant, N. et al. Identification and estimation of bioactive constituents Negundoside, Berberine chloride, and Marmelosin by HPLC and HPTLC for development of quality control protocols for Ayurvedic medicated oil formulation. Future Journal of Pharmaceutical Sciences. 2021;7(1):171. https://doi.org/10.1186/s43094-021-00322-3CrossrefGOOGLE SCHOLAR
  23. 23.Mo, Y.; Ma, J.; Gao, W.; Zhang, L.; Li, J.; Li, J. et al. Pomegranate Peel as a Source of Bioactive Compounds: A Mini Review on Their Physiological Functions. Frontiers in Nutrition. 2022;9.GOOGLE SCHOLAR
  24. 24.Mohan, M.; A, M. C.; D, P.; V, A. G.. Review of Pharmacological and Medicinal Uses of Punica granatum. Cureus. 2024;16(10). https://doi.org/10.7759/cureus.71510CrossrefGOOGLE SCHOLAR
  25. 25.Mokgalaboni, K.; Dlamini, S.; Phoswa, W. N.; Modjadji, P.; Lebelo, S. L.. The Impact of Punica granatum Linn and Its Derivatives on Oxidative Stress, Inflammation, and Endothelial Function in Diabetes Mellitus: Evidence from Preclinical and Clinical Studies. Antioxidants. 2023;12(8). https://doi.org/10.3390/antiox12081566CrossrefGOOGLE SCHOLAR
  26. 26.Nuncio-Jáuregui, N.; Munera-Picazo, S.; Calín-Sánchez, Á.; Wojdyło, A.; Hernández, F.; Carbonell-Barrachina, Á. A.. Bioactive compound composition of pomegranate fruits removed during thinning. Journal of Food Composition and Analysis. 2015;37:11–19. https://doi.org/10.1016/j.jfca.2014.06.015CrossrefGOOGLE SCHOLAR
  27. 27.Popovici, V.; Bucur, L.; Calcan, S. I.; Cucolea, E. I. et al. Elemental Analysis and In vitro Evaluation of Antibacterial and Antifungal Activities of Usnea barbata (L.) Weber ex F.H. Wigg from Călimani Mountains, Romania. Plants. 2021;11(1):32. https://doi.org/10.3390/plants11010032CrossrefGOOGLE SCHOLAR
  28. 28.Ramlagan, P.; Labib, R. M.; Farag, M. A.; Neergheen, V. S.. Advances towards the analysis, metabolism and health benefits of punicalagin, one of the largest ellagitannin from plants, with future perspectives. Phytomedicine Plus. 2022;2(3):100313. https://doi.org/10.1016/j.phyplu.2022.100313CrossrefGOOGLE SCHOLAR
  29. 29.Ranjha, M. M. A. N.; Shafique, B.; Wang, L. et al. A comprehensive review on phytochemistry, bioactivity and medicinal value of bioactive compounds of pomegranate (Punica granatum). Advances in Traditional Medicine. 2023;23(1):37–57. https://doi.org/10.1007/s13596-021-00566-7CrossrefGOOGLE SCHOLAR
  30. 30.Riolo, M.; Luz, C.; Bua, C.; Barreca, S.; Tambè, M. C. et al. Qualitative and relative abundance analysis of Alternaria alternata metabolites in pomegranate (Punica granatum L.) cultivars affected by Heart Rot using UHPLC-Q-TOF-MS. Food Control. 2025;172. https://doi.org/10.1016/j.foodcont.2025.111204CrossrefGOOGLE SCHOLAR
  31. 31.Shabir, I.; Dar, A. H.; Dash, K. K.; Manzoor, S. et al. Bioactive potential of punicalagin: A comprehensive review. Applied Food Research. 2024;4(2). https://doi.org/10.1016/j.afres.2024.100572CrossrefGOOGLE SCHOLAR
  32. 32.Shahkoomahally, S.; Shin, D.; Habibi, F.; Kim, J.; Sarkhosh, A.. Profiling phenolic compounds in juice and peel of fourteen pomegranate (Punica granatum L.) varieties grown in Florida, USA. Food Chemistry Advances. 2023;2. https://doi.org/10.1016/j.focha.2023.100225CrossrefGOOGLE SCHOLAR
  33. 33.Siddiqui, S. A.; Singh, S.; Nayik, G. A.. Bioactive compounds from pomegranate peels - Biological properties, structure-function relationships, health benefits and food applications - A comprehensive review. Journal of Functional Foods. 2024;116. https://doi.org/10.1016/j.jff.2024.106132CrossrefGOOGLE SCHOLAR
  34. 34.Singh, B.; Singh, J. P.; Kaur, A.; Singh, N.. Phenolic compounds as beneficial phytochemicals in pomegranate (Punica granatum L.) peel: A review. Food Chemistry. 2018;261:75–86. https://doi.org/10.1016/j.foodchem.2018.04.039CrossrefGOOGLE SCHOLAR
  35. 35.Skrt, M.; Albreht, A.; Vovk, I.; Constantin, O. E.; Râpeanu, G. et al. Extraction of Polyphenols and Valorization of Fibers from Istrian-Grown Pomegranate (Punica granatum L.). Foods. 2022;11(18):2740. https://doi.org/10.3390/foods11182740CrossrefGOOGLE SCHOLAR
  36. 36.Tang, L.; Li, T.; Zhang, B.; Zhang, Z.; Sun, X.. Punicalagin Alleviates Psoriasis by Inhibiting NF-κB-Mediated IL-1β Transcription and Caspase-1-Regulated IL-1β Secretion. Frontiers in Pharmacology. 2022;13. https://doi.org/10.3389/fphar.2022.817526CrossrefGOOGLE SCHOLAR
  37. 37.Venkatesham, B.; Chaithra, D.; Naikodi, M. A. R.; Nazeer, M.; Siddiqui, A.. Pharmacognostic evaluation, physicochemical standardization and HPTLC fingerprint analysis of pomegranate (Punica granatum L.) leaf and seed. Annals of Phytomedicine: An International Journal. 2021;10(2):187–194. https://doi.org/10.21276/ap.2021.10.2.26CrossrefGOOGLE SCHOLAR
  38. 38.Venusova, E.; Kolesarova, A.; Horky, P.; Slama, P.. Physiological and Immune Functions of Punicalagin. Nutrients. 2021;13(7):2150. https://doi.org/10.3390/nu13072150CrossrefGOOGLE SCHOLAR
  39. 39.Wang, W.; Long, P.; He, M.; Luo, T.; Li, Y. et al. Pomegranate polyphenol punicalagin as a nutraceutical for mitigating mild cognitive impairment: An overview of beneficial properties. European Journal of Pharmacology. 2024;977. https://doi.org/10.1016/j.ejphar.2024.176750CrossrefGOOGLE SCHOLAR
  40. 40.Xu, J.; Cao, K.; Liu, X.; Zhao, L.; Feng, Z.; Liu, J.. Punicalagin Regulates Signaling Pathways in Inflammation-Associated Chronic Diseases. Antioxidants. 2021;11(1). https://doi.org/10.3390/antiox11010029CrossrefGOOGLE SCHOLAR
  41. 41.Yisimayili, Z.; Chao, Z.. A review on phytochemicals, metabolic profiles and pharmacokinetics studies of the different parts (juice, seeds, peel, flowers, leaves and bark) of pomegranate (Punica granatum L.). Food Chemistry. 2022;395(5). https://doi.org/10.1016/j.foodchem.2022.133600CrossrefGOOGLE SCHOLAR

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Chowdareddy, R. P., Jayaraman, T., AK, A. K., Ghanta, M., & Umeza, B. (2027). Comprehensive HPTLC Fingerprinting and Quality Evaluation of Punicalagin in Pomegranate (Punica granatum L.) Aril Juice. International Journal of Pharmaceutical Investigation, 17(1), 249–255. https://doi.org/10.5530/ijpi.20270024