A Novel Densitometric HPTLC Method Integrated with FTIR for The Detection of Rutin in Senna auriculata Flower Extract

A Novel Densitometric HPTLC Method Integrated with FTIR for The Detection of Rutin in Senna auriculata Flower Extract K. Deepashree Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology & Advanced Studies, Pallavaram, Chennai, Tamilnadu, India https://orcid.org/0009-0002-0066-0570 M. Sumithra Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology & Advanced Studies, Pallavaram, Chennai, Tamilnadu, India https://orcid.org/0000-0003-3781-107X P. Indhumathy Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology & Advanced Studies, Pallavaram, Chennai, Tamilnadu, India https://orcid.org/0000-0003-4080-7178 N. Narendra Kumar Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology & Advanced Studies, Pallavaram, Chennai, Tamilnadu, India https://orcid.org/0009-0007-4775-6736 M. Archana Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology & Advanced Studies, Pallavaram, Chennai, Tamilnadu, India https://orcid.org/0000-0001-7264-2182

The objective of this research was to create a reliable way to analyze flavonoids, mainly Rutin, in extract from the Senna auriculata flower with densitometric HPTLC and FTIR. Fresh flowers were extracted using methanol and analyzed on silica gel plates using toluene\:ethyl acetate\:formic acid as the mobile phase, looking for compounds with toluene (254 nm) and ethyl acetate (366 nm). Rutin was confirmed in the Standard Rutin and the extract, as the Rf values for both were 0.014, 0.017 for Standard Rutin and 0.032, 0.026 for the extract. There was 1.12% Rutin present in the raw material. The presence of phenolic, aromatic and ether functional groups in FTIR confirmed that the compounds are flavonoids. Using this approach, the process was reliable, effective, pollution-free and needed very little solvent. The method enables the generation of a radiation-proof, non-rinse herbal sheet mask for space missions, specifically those on the International Space Station (ISS).
10 31 2025 10 31 2025 1518 1527 https://doi.org/10.13003/0pyd3js 2025-10-31 http://creativecommons.org/licenses/by/4.0/ 10.13005/ojc/410508 https://www.orientjchem.org/vol41no5/a-novel-densitometric-hptlc-method-integrated-with-ftir-for-the-detection-of-rutin-in-senna-auriculata-flower-extract/ https://www.orientjchem.org/vol41no5/a-novel-densitometric-hptlc-method-integrated-with-ftir-for-the-detection-of-rutin-in-senna-auriculata-flower-extract/ 10.1007/s11120-009-9439-x 1. Berthomieu, C., Hienerwadel, R. Fourier transform infrared (FTIR) spectroscopy. Res., 2009, 101(2–3), 157–170. 2. Kumar, A., et al. Estimation of gallic acid, rutin, and quercetin in Terminalia chebula by HPTLC. Jordan J. Pharm. Sci., 2010, 3(1), 63–68. 10.1556/jpc.23.2010.1.7 3. Sun, J., et al. Simultaneous HPTLC analysis of flavonoids in the leaves of three different species of bamboo. Planar Chromatogr., 2010, 23(1), 40–45. 4. Ahmad, H., et al. Determination of quercetin in Michelia champaca (Champa) leaves and stem bark by HPTLC. Int. J. Pharma Bio Sci., 2011. 5. Dhandapani, A., Kadarkarai, M. HPTLC quantification of flavonoids, larvicidal and smoke repellent activities of Cassia occidentalis (Caesalpiniaceae). J. Phytol., 2011, 3(2), 60–72. 6. Ananth, P.H., et al. Development of validated HPTLC method for simultaneous quantification of rutin and quercetin. J. Pharm., 2012, 2(1), 33–38. 10.1016/S2221-1691(11)60112-3 7. Mariswamy, Y., et al. Chromatographic fingerprint analysis on flavonoid constituents of Aerva lanata Asian Pac. J. Trop. Biomed., 2012, 2(Suppl 1), S8–S12. 10.1016/j.phme.2013.12.004 8. Amir, M., et al. Development and validation of HPTLC method for quercetin and rutin. Methods, 2013, 4(1), 62–67. 9. Rao, A.S., et al. Simultaneous estimation of quercetin and rutin. Asian J. Biomed. Pharm. Sci., 2013, 3(21), 56–59. 10. Gowda, J.S., Veerabhadrappa, S.B. In vitro antioxidant activity and HPTLC fingerprint of quercetin in Cassia auriculata. Asian J. Plant Sci. Res., 2013, 3(4), 162–169. 11. Bhalke, R.D., et al. Estimation of rutin, quercetin, and gallic acid in Pterospermum acerifolium. Indo Am. J. Pharm. Res., 2013, 3(9), 7008–7017. 10.1002/jssc.200700066 12. Bhandari, P., et al. RP-HPTLC method for simultaneous determination of major flavonoids. Sep. Sci., 2007, 30(14), 2092–2096. 13. Ananthi, T., Keerthana, R. HPTLC analysis: Identification of flavonoid from Senna auriculata Int. J. Sci. Res. Med., 2016, 3(3), 64–72. 10.1365/s10337-006-0822-x 14. Abou-Donia, A.H., et al. Determination of rutin in Amaryllis belladonna. Chromatographia, 2006, 64(1–2), 109–112. 10.1556/JPC.18.2005.4.3 15. Bilušić Vundać, V., et al. Flavonoids and phenolic acids in Croatian Stachys J. Planar Chromatogr., 2005, 18(4), 269–273. 10.4314/ijmu.v2i2.39851 16. Lakhanpal, P., Rai, D.K. Quercetin: A versatile flavonoid. J. Med. Update, 2007, 2, 22–37. 10.1080/10826070701451621 17. Hawrył, M.A., et al. Separation of flavonoids by RP-HPLC/NP-HPTLC. Liq. Chromatogr. Relat. Technol., 2007, 30(15), 2253–2265. 10.1177/1934578X1200700918 18. Avula, B., et al. Simultaneous determination of alkaloids and flavonoids. Prod. Commun., 2012, 7(9), 1177–1180. 19. Bazylko, A., et al. Flavonoids in Galinsoga parviflora and ciliata. Phytochem. Lett., 2014, 10, 179–185. 10.1080/10942912.2014.988722 20. Ahil, S.B., et al. Comparative HPTLC analysis of safflower cultivars. J. Food Prop., 2015, 18(11), 2561–2570. 21. Patil, V., et al. Simultaneous estimation of rutin, quercetin, and liquiritin in Cocculus hirsutus. J. Pharmacogn., 2015, 2(4), 49–55. 22. Arunachalam, K.D., et al. HPTLC and phytochemical analysis of Morinda tinctoria. J. Pharm. Pharm. Sci., 2015, 7(2), 360–366. 10.4103/0974-8490.171098 23. Doshi, G.M., Une, H.D. Quantification of quercetin and rutin. Res., 2016, 8(1), 37–42. 24. Omosa, L.K., et al. Surface exudates and antioxidant activity of Dodonaea angustifolia. Nat. Sci. Res., 2016, 6(6). 10.4103/0973-7847.194044 25. Anand David, A.V., et al. Biological importance of quercetin. Rev., 2016, 10(20), 84–89. 10.1002/pca.2621 26. Kroslakova, I., et al. Coupling of HPTLC with MALDI-TOF MS. Anal., 2016, 27(3), 222–228. 10.1556/jpc.22.2009.1.5 27. Bros, I., et al. HPTLC quantification of flavonoids in Satureja hortensis. Planar Chromatogr., 2009, 22(1), 25–28. 10.1080/13880209.2017.1300175 28. Alam, P., et al. Quantitative analysis of rutin, quercetin, naringenin. Biol., 2017, 55(1). 10.1016/j.biopha.2018.10.007 29. Gayathri, N., et al. Antidiabetic activity of Cassia auriculata. Pharmacother., 2018, 108, 1495–1506. 10.1007/s12161-018-1332-9 30. Bajkacz, S., et al. Determination of flavonoids and phenolic acids. Food Anal. Methods, 2018, 11, 3563–3575. 10.1007/s00764-020-00035-y 31. Satija, S., et al. HPTLC fingerprint for berberine and rutin. Planar Chromatogr., 2020, 33(5), 435–443. 10.1002/pca.2749 32. Shawky, E., Abou El Kheir, R.M. Discrimination of Apiaceae species. Anal., 2018, 29(6), 675–685. 10.18203/2319-2003.ijbcp20175692 33. Doshi, G.M., et al. Rutin and quercetin quantification and anticancer activities. J. Basic Clin. Pharmacol., 2018, 7(1), 153–161. 34. Veerachari, U., et al. Chemical profiling of Cassia Eur. J. Biotechnol. Biosci., 2019, 7(4), 50–59. 35. Orsini, F., et al. HPTLC-MS/MS and antioxidant activity in Cyclanthera pedata. Liq. Chromatogr. Relat. Technol., 2019, 42(18–19), 498–506. 10.5958/0974-360X.2020.00484.9 36. Kharat, S.N., et al. Flavonoid screening in Syzygium cumini. J. Pharm. Technol., 2020, 13(6), 2720–2726. 10.1177/1934578X21991723 37. Agrawal, P.K., et al. Rutin as potential SARS-CoV-2 Mpro inhibitor. Prod. Commun., 2021, 16(4). 10.3389/fphar.2022.1000608 38. Dehelean, C.A., et al. Rutin bioconjugates. Pharmacol., 2022, 13, 1000608. 10.1002/ptr.6904 39. Negahdari, R., et al. Therapeutic benefits of rutin. Res., 2021, 35(4), 1719–1738. 10.1016/j.sajb.2020.11.017 40. Oladeji, O.S., et al. Review of Senna. Afr. J. Bot., 2021, 138, 1–32. 10.3390/plants11091117 41. Drețcanu, G., et al. Role of flavonoids in cancer prevention. Plants (Basel), 2022, 11(9), 1117. 10.3390/plants10030475 42. Rodríguez-Valdovinos, K.Y., et al. Quantitative analysis of rutin. Plants, 2021, 10(3), 475. 10.1155/2014/724267 43. Chelyn, J.L., et al. Flavone C-glycosides in Clinacanthus nutans. World J., 2014, 724267. 10.3390/molecules28062870 44. Tessema, F.B., et al. HPTLC profile of Dodonaea angustifolia. Molecules, 2023, 28(6), 2870. 10.4103/jpbs.jpbs_551_23 45. Rahman, A.F.K., et al. Antimicrobial activity of Senna auriculata. Pharm. Bioallied Sci., 2024, 16(Suppl 2), S1233–S1236. 46. Thakkar, M., Sharma, S. HPTLC of flavonoids in polyherbal formulation. J. Pharm. Sci. Res., 2023, 14(2), 876–882. 47. Mohanakumaran, K., et al. Detection of flavonoids by HPTLC. World J. Pharm. Sci., 2025, 13(1), 90–94. 48. Huria, N., et al. Estimation of rutin in Morus Int. J. Pharm. Sci., 2024, 2(3), 565–577. 10.5530/pres.15.3.058 49. Radha, P., et al. HPTLC profiling of Senna Pharmacogn. Res., 2023, 15(3), 551–561. 10.1016/j.arabjc.2021.103345 50. Prasathkumar, M., et al. Bioactivity of Senna auriculata Arab. J. Chem., 2021, 14(9), 103345. 10.1016/j.heliyon.2022.e10103 51. Bationo, R.K., et al. Flavonoids of Cymbopogon giganteus. Heliyon, 2022, 8, e10103. 10.1186/s43094-023-00571-4 52. Ansari, H.I., et al. New flavonoid from Abrus precatorius. Future J. Pharm. Sci., 2023, 9, 119. 10.33263/LIANBS113.38613870 53. Jain, D., et al. TLC and HPTLC fingerprinting of Cyperus rotundus. Appl. NanoBioSci., 2022, 11(3), 3861–3870. 10.3390/molecules26226892 54. Shawky, E., et al. FTIR fingerprinting of olive leaf extracts. Molecules, 2021, 26, 6892. 10.1155/2017/8647212 55. Ye, Z., et al. Chemical differentiation in Dendrobium. Based Complement. Alternat. Med., 2017, 8647212. 56. Hawrył, M.A., et al. Rutin determination in Benincasa hispida. Chromatogr., 2021, e5230. 57. Kao Corporation. Kao’s technology for skin health. Press Release, 2021 Nov 22. 10.1080/13880209.2017.1300175 58. Alam, P., et al. RP- and NP-HPTLC for anti-HBV extracts. Biol., 2017, 55(1). 10.3390/molecules26226892 59. Agatonovic-Kustrin, S., et al. FTIR characterization of flavonoids. Molecules, 2021, 26. 10.1002/pca.2749 60. Shawky, E., et al. Multivariate image analysis of Apiaceae Phytochem. Anal., 2018, 29(6), 675–685.