Urolithiasis: HPTLC Method for Quantitative Detection of Rutin and Quercetin in an Herbal Plant


  • Patel Ravindrakumar Department of Pharmacology, Indukaka Ipcowala College of Pharmacy, New Vallabh Vidyanagar, Anand - 388121, Gujarat
  • Niraj Vyas Department of Pharmacognosy and Phytochemistry, Ramanbhai Patel College of Pharmacy, Changa, Anand - 388421, Gujarat
  • Patel Sandip Department of Pharmacology, L.M. College of Pharmacy, Ahmedabad - 380058, Gujarat




Densitometry, <i>Ocimum basilicum</i>, Quercetin, High-performance Thin-layer Chromatography (HPTLC), Rutin, Validation


Anti-inflammatory, Hypoglycemic, Hepatoprotective, Antihyperlipidemic, Anti-Ulcerative, Cardioprotective Stimulant, Sedative, Hypnotic, Anticonvulsant Activity, Memory Retention, Stroke Preventive Activity, Antimicrobial Activity, Antimycobacterial, Antiviral, Larvicidal, Antiparasitic, Chemopreventative, Chemo-modulatory, Anticancer, Cytoprotective activities were found in a variety of herbal plants, including Ocimum basilicum. The purpose of this study was to design and develop a new HPTLC method that was accurate, precise and cost-effective for simultaneous measurement of rutin and quercetin in a hydroalcoholic extract of Ocimum basilicum seeds. The mobile phase was Toluene: Ethyl Acetate: Methanol: formic acid (6:4:3:1, v/v/v/v/v) and densitometric scanning was performed at 254 nm. Merck TLC aluminium sheets of silica gel 60 F254, (10 x 10 cm) with a thickness of 250 mm was used as stationary phase. At a wavelength of 254 nm, rutin and quercetin were detected. The constituents were resolved satisfactorily, with Rf values of 0.25 ± 2.01 for rutin and 0.80 ± 0.64 for quercetin, respectively. Linearity (300-1300 ng/spot for rutin and quercetin) was used to test the method’s accuracy and reproducibility. For both analytes, intra- and inter-day precision, as evaluated by coefficient of variation, was less than 3%. For rutin, the detection and quantification limits were 46.52 and 140.96 ng/spot, respectively, and for quercetin, they were 81.79 and 247.84 ng/spot. The proposed approach was found to be precise, accurate, repeatable, and specific, and it could be used to measure quercetin and rutin in samples at the same time.


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Schreiner M, Huyskens-Keil S. Phytochemicals in fruit and vegetables: Health promotion and postharvest elicitors. Cri Rev Plant Sci. 2006; 25:267-78. https://doi.org/10.1080/07352680600671661

Soleas GJ, Grass L, Josephy PD, Goldberg DM, Diamandis EP. A comparison of the anticarcinogenic properties of four red wine polyphenols. Clin Biochem. 2002; 35:119-24. https://doi.org/10.1016/S0009-9120(02)00275-8

Middleton EM, Teramura AH. The role of flavonol glycosides and carotenoids in protecting soybean from ultraviolet-B damage. Plant Physiol. 1993; 103:741-52. https://doi.org/10.1104/pp.103.3.741. PMid:12231976. PMCid:PMC159044

Hein KE, Tagliaferro AR, Bobilya DJ. Flavonoid antioxidants: Chemistry, metabolism and structure-activity relationships. J Nutr Biochem. 2002; 13(10):572-84. https://doi.org/10.1016/S0955-2863(02)00208-5

Bors W, Heller W, Michel C, Saran M. Flavonoids as antioxidants: determination of radical-scavenging efficiencies. Method Enzymol. 1990; 186:343-55. https://doi.org/10.1016/0076-6879(90)86128-I

Dixon RA, Dey PM, Lamb CJ. Phytoalexins: enzymology and molecular biology. Adv Enzymol RAMB.1983; 55:1-136.

Rice-Evans CA, Miller NJ, Bolwell PG, Broamley PM, Pridham JB. The relative antioxidant activities of plantderived polyphenolic flavonoids. Free Radical Res. 1995; 22(4):375-83. https://doi.org/10.3109/10715769509145649. PMid:7633567

Agati G, Azzarello E, Pollastri S, Tattini M. Flavonoids as antioxidants in plants: location and functional significance. Plant Sci. 2012; 196:67-76. https://doi.org/10.1016/j.plantsci.2012.07.014. PMid:23017900

Du F, Zhang F, Chen F, Wang A. Advances in microbial heterologous production of flavonoids. Afr J Microbiology Res. 2011; 5(18):2566-74. https://doi.org/10.5897/AJMR11.394

Middleton EJ. Effect of plant flavonoids on immune and inflammatory cell function. Adv Exp Med Biol. 1998; 439:175-82. https://doi.org/10.1007/978-1-4615-5335-9_13. PMid:9781303

Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. Scientific World J. 2013; 2013:1- 16. https://doi.org/10.1155/2013/162750. PMid:24327805. PMCid:PMC3845396

Nijveldt RJ, Nood EL, Van Horn DEC, Boelens PG, Van Norren K, van Leeuwen PAM. Flavonoids: A review of probable mechanisms of action and potential applications. Am J Clin Nutr. 2001; 74(4):418-25. https://doi.org/10.1093/ajcn/74.4.418. PMid:11566638

Bilal A, Jahan N, Ahmed A, Bilal SN, Habib S, Hajra S. Phytochemical and pharmacological studies on Ocimum basilicum linn — A review. Int J Curr Res. 2012; 4(23).

International Conference on Harmonization. Validation of Analytical Procedures Methodology Q2B; 1996.

Method for determination of forskolin in crude drug and pharmaceutical dosage form. Chromatographia. 2008; 67:441-47. https://doi.org/10.1365/s10337-008-0521-x

Alam P, Ali M, Ahmad SA. A validated HPLC method for estimation of cordifolioside A in Tinospora cardifolia, Miers and marketed formulations. J Chromatogr Sci. 2009; 47: 910-13. https://doi.org/10.1093/chromsci/47.10.910. PMid:19930804

Parveen R, Baboota S, Ahmad S, Ali J, Ahuja A. Stabilityindicating HPTLC method for quantitative estimation of silybin in bulk drug and pharmaceutical dosage form. Biomed Chromatogr. 2010; 24:639-47. https://doi.org/10.1002/bmc.1340. PMid:19816854

Rozylo JK, Janicka M. Different planar techniques for prediction of solute retention in column liquid chromatography. J Planar Chromatogr. 1996; 9:418-24.

Weins C, Hauck HE. Advances and developments in thin layer chromatography. LC-GC Int. 1996; 4:455-71.

Poole CF, Poole SK. Instrumental thin layer chromatography. Anal Chem. 1994; 66:27A-37A. https://doi.org/10.1021/ac00073a001

Sejal P, Niraj V. Validated spectrofluorimetric method for estimation of piperine in an ayurvedic formulation. Asian J Pharm Clin Res. 2012; 5(4):231-233.

Niraj V, Sangita P. Development and Validation of RP-HPLC Method for Simultaneous estimation of Nebivolol and Indapamide in Pharmaceutical Dosage Form. Asian J Pharm Anal. 2014; 4(3):98-102.




How to Cite

Ravindrakumar, P., Vyas, N., & Sandip, P. (2022). Urolithiasis: HPTLC Method for Quantitative Detection of Rutin and Quercetin in an Herbal Plant. Journal of Natural Remedies, 22(3), 371–379. https://doi.org/10.18311/jnr/2022/29339



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