HPTLC Fingerprinting and Validation of Quercetin Yield of Gel Formulation for Diabetic Neuropathic Pain

Authors

  • Ankita Raikwar
  • Mahesh Kumar Posa
  • Subhranshu Panda
  • Md. Shamsuzzman

DOI:

https://doi.org/10.63682/jns.v14i32S.9869

Keywords:

HPTLC, Quercetin, Flavonoid, Poly herbal, Gel

Abstract

Background: The growing global dependence on herbal medicines highlights the urgent need for validated analytical techniques to ensure their quality, safety, and therapeutic efficacy. This study aimed to develop and validate a High-Performance Thin Layer Chromatography (HPTLC) method for the quantitative estimation of quercetin, a potent antioxidant flavonoid, in a polyherbal formulation.

Material and Methods: HPTLC method was developed using a mobile phase composed of toluene: ethyl acetate (9:1 v/v), which provided a sharp, well-resolved peak for quercetin at an Rf value of 0.892. Method validation was performed according to ICH Q2 (R1) guidelines, evaluating specificity, linearity, accuracy, precision, robustness, and sensitivity parameters.

Results: The method showed excellent linearity over the concentration range of 1000–5000 ng/spot, with a correlation coefficient (r²) of 0.999. The limit of detection (LOD) and limit of quantification (LOQ) were determined as 60.67 ng/spot and 174.00 ng/spot, respectively. Accuracy was confirmed through recovery studies, yielding results between 99.12% and 99.85%. Intra-day precision exhibited %RSD values below 2%, demonstrating high repeatability. Robustness tests indicated the method’s resilience to slight variations in experimental conditions. The quercetin content in the gel formulation was found to be approximately 13.76 mg/g.

Conclusion: The developed HPTLC method is simple, accurate, and reliable for the quantitative estimation of quercetin in polyherbal formulations. It is suitable for routine quality control and standardization, supporting the safety and efficacy of herbal medicinal products. ..

Downloads

Download data is not yet available.

References

Panche AN, Diwan AD, Chandra SR. Flavonoids: an overview. J. Nutritional Sci. 2016; 5:47.

2. Wenzel U. Flavonoids as drugs at the small intestinal level. Current opinion in Pharmacol. 2013; 1,13(6):864-8.

3. Safe S, Jayaraman A, Chapkin RS, Howard M, Mohankumar K, Shrestha R. Flavonoids: structure–function and mechanisms of action and opportunities for drug development. Toxicolo. Res. 2021; 37(2):147-62.

4. Rana, A. C., & Gulliya, B. Chem. and pharmaco. of flavonoids-A review. Indian J. Pharmaceut. Edu. Res., 2019 53(1).

5. Mohamed, T. A et al., Plant cell cultures: An enzymatic tool for polyphenolic and flavonoid transformations. Phytomedicine, 2022 100, 154019.

6. Aghababaei F, Hadidi M. Recent advances in potential health benefits of quercetin. Pharmaceu. 2023; 16(7):1020.

7. Al Mamun A, et al., Exploring the role of senescence inducers and senotherapeutics as targets for anticancer natural products. Euro. J. Pharmacol. 2022; 928 :174991.

8. Teodoro JS, Machado IF, Castela AC, Rolo AP, Palmeira CM. Mitochondria as a target for safety and toxicity evaluation of nutraceuticals. In Nutraceuticals. 2021; 1, 463-483. Academic Press.

9. Panchal H, Amin A, Shah M. Development of validated high-performance thin-layer chromatography method for simultaneous determination of quercetin and kaempferol in Thespesia populnea. Pharmacog. Res. 2017; 9(3):277.

10. Doshi GM, Une HD. Quantification of quercetin and rutin from Benincasa hispida seeds and Carissa congesta roots by high-performance thin layer chromatography and high-performance liquid chromatography. Pharmacogn. Res. 2016; 8(1):37.

11. Das B, Saha P, Maity N. Quantitative Estimation of Quercetin of Some Selected Edible Plants of West Bengal by High-performance Thin-layer Chromatography Densitometry Method. Asian J. Pharmaceut. Res. Health Care. 2023; 1;15(3):254-61.

12. Verma A, Patel P, Sharma A, Kurmi BD. Quercetin in ayurvedic formulations: High- performance thin-layer chromatography based rapid fingerprinting profiling. Phytomed. Plus. 2023; 3(1):100416.

13. Shah MS, Dama GY. Development and validation of high-performance thin layer

chromatography method for simultaneous determination of gallic acid and quercetin from Annona squamosa L. leaf extract. Res. J. Pharmacy Techno. 2024; 17(3):1200-6.

14. Patel AA, Amin AA, Patwari AH, Shah MB. Validated high performance thin layer chromatography method for simultaneous determination of quercetin and gallic acid in Leea indica. Revista Brasileira de Farmacognosia. 2017; 27(1):50-3.

15. Vyas A, Jain V, Sahu U, Kumar N, Joshi N. HPTLC method development of herbal drugs and its validation: An overview. Res. J. Pharmacy Tech. 2023; 16(8):3964-76.

16. Guideline IH. Validation of analytical procedures Q2 (R2). ICH: Geneva, Switzerland. 2022; 1.

17. Balap AR, Dudhe AP. Stability Indicating HPTLC Method Development and Validation for Quantification of Favipiravir in Bulk and Commercial Film Coated Tablet. Inter. J. Pharmaceut. Investigation. 2023; 1;13(3).

18. Lodha SR, Patel KD, Patel SA, Patel SG, Bodiwala KB, Shah SA, Kalyankar GG. Development and validation of HPTLC method for estimation of teneligliptin hydrobromide hydrate in tablet dosage form. J Pharm Appl Sci. 2016; 3(1):27-34.

19. Gandhi SP, Dewani MG, Borole TC, Damle MC. Development and validation of stability indicating HPTLC method for determination of Ofloxacin and Ketorolac Tromethamine in combination. J. Advanced Scientific Res. 2011; 10; 2(03):77-82.

20. Shah P, Patel J, Patel K, Gandhi T. Development and validation of an HPTLC method for the simultaneous estimation of Clonazepam and Paroxetine hydrochloride using a DOE approach. J. Taibah Uni. Sci. 2017;11(1):121-32.

Downloads

Published

2025-10-18

How to Cite

1.
Raikwar A, Kumar Posa M, Panda S, Shamsuzzman M. HPTLC Fingerprinting and Validation of Quercetin Yield of Gel Formulation for Diabetic Neuropathic Pain. J Neonatal Surg [Internet]. 2025 Oct. 18 [cited 2026 Apr. 14];14(32S):10295-301. Available from: https://jneonatalsurg.com/index.php/jns/article/view/9869

Issue

Vol. 14 No. 32S (2025): Journal of Neonatal Surgery

Section

Original Article

License

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

You are free to:

  • Adapt — remix, transform, and build upon the material for any purpose, even commercially.

Terms:

  • Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
  • No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.