A NOVEL, SIMPLE, RAPID RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF FERULIC ACID, QUERCETIN, PIPERINE AND THYMOL IN AYURVEDIC FORMULATION
Objective: A simple, accurate, precise and robust reverse phase high performance liquid chromatography (RP-HPLC) method was developed for simultaneous estimation of ferulic acid, quercetin, piperine and thymol in marketed ayurvedic formulation.
Methods: The selected markers were resolved using shim pack GIST C-18 column, with mobile phase acetonitrile: 0.04 M potassium dihydrogen ortho phosphate buffer (pH 3.0 adjusted with ortho phosphoric acid) in a ratio of 60:40 v/v at a flow rate of 1.0 ml/min. The detection was carried out at 264 nm.
Results: The retention time of ferulic acid, quercetin, piperine and thymol were found to be 2.98, 3.35, 7.83 and 9.72 min respectively. The developed method was validated according to the guidelines provided in ICH Q2 (R1) in term of linearity, precision, limit of detection, limit of quantification, accuracy and robustness. Linear response for all selected markers was obtained in the concentration range of 12-28 Âµg/ml with a correlation coefficient (r2) greater than 0.999. The mean % recovery was found to be 99.30 for ferulic acid, 98.77 for quercetin, 100.93 for piperine and 100.25 for thymol.
Conclusion: The developed method was applied for quantification of these marker in marketed ayurvedic formulation. This method can be used to evaluate other formulations containing these selected phytoconstituent, thus conforming the quality and safety of ayurvedic or polyherbal formulations.
2. Meena MC, Patni V. Isolation and identification of flavonoid quercetin from Citrullus colocynthis (Linn.) Schrad. AJES 2008;22:137-42.
3. Jadhav AP, Kareparamban JA, Nikam PH, Kadam VJ. A validation high-performance liquid chromatography method for estimation of ferulic acid in asafoetida and polyherbal preparation. Indian J Pharm Sci 2013;75:493-5.
4. Baskaran N, Manoharan S, Balakrishnan S. Chemopreventive potential of ferulic acid in 7,12-dimethylbenz[a] anthracene-induced mammary carcinogenesisin sprague-dawley rats. Eur J Pharmacol 2010;637:22-9.
5. Cheng CY, Su SY, Tang NY. Ferulic acid provides neuroprotection against oxidative stress-related apoptosis after cerebral ischemia/perfusion injury by inhibiting ICAM-1 mRNA expression in rats. Brain Res 2008;2:90.
6. Middleton E, Kandaswami C. The impact of plant flavonoids on mammalian biology: implications for immunity, inflammation and cancer, in the flavonoids. advances in research science harborne, I. R. London: Chapman and Hall; 1993. p. 619-45.
7. Jain V, Shaikh S. Simulaneous RP-HPLC analysis of quercetin and kaempferol in different plant part of Cissus quadrangularis. Int J Pharm Pharm Sci 2016;8:138-42.
8. Jain V, Shaikh SGM. Development and validation of a RP-HPLC method for the simultaneous determination of curcumin, piperine and camphor in an ayurvedic formulation. Int J Pharm Pharm Sci 2018;10:115-21.
9. Kapuriya KG, Parmar PM, Topiya HR, Faldu SD. Method development and validation of rifampicin and piperine in their combined dosage form. Int Bull Drug Res 2012;01:71-80.
10. Shah U, Jasani A. UV spectrophotometric and RP-HPLC methods for simultaneous estimation of isoniazid, rifampicine and piperine in pharmaceutical dosage form. Int J Pharm Pharm Sci 2014;6:274-80.
11. Rustaiyan A, Lajevardi T, Rabbani M, Yari M, Masoudi S. Chemical constituents of the essential oil of Thymus kotschyanus Boiss. and Hohen. Iran Daru 1999;7:27-8.
12. Hajimehdipoor H, Khanavi M, Zahedi H, Abedi Z. Fingerprint study of thymus spp. by TLC. J Med Plants 2009;8:19-24.
13. International Conference on Harmonization of Technical Requirement for Registration of Pharmaceuticals for Human use. Validation of Analytical Procedure: Text and Methodology ICH Q2 (R1); 2005.
14. Vemula VR, Sharma PK. RP-HPLC method development and validation for simultaneous estimation of diclofenac and tolperisone in tablet dosage form. Asian J Pharm Clin Res 2013;6 Suppl 3:186-9.
15. Shah R, Shah R. Development and validation of an RP-HPLC method for phenytoin sodium and phenobarbitone in bulk and pharmaceutical dosage form. Int J Pharm Pharm Sci 2017;9:224-9.
16. FDA, Guidance R. Validation of chromatographic methods. Center for drug evaluation and research (CDER), Food Drug Administration; 1994. p. 2.
17. FDA, ORA validation and verification guidance for human drug analytical methods. Food and Drug Administration; 2003. p. 1.