DEVELOPMENT AND VALIDATION OF NOVEL REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF ANDROGRAPHOLIDE AND ALOE-EMODIN
Objective: The study aims to develop and validate a novel reverse-phase high-performance liquid chromatographic method for simultaneous estimation of andrographolide and aloe-emodin in herbal formulation and validate as per the International Conference on Harmonization (ICH) guidelines.
Methods: The analysis was carried on a Shimadzu LC Prominence-i 2030 model with the Lab Solution software. The column used for separation was Prontosil C18 (250×4.6 mm, 5 μ), with a mobile phase consisting of acetonitrile and 0.05% orthophosphoric acid (45:55), at a flow rate of 1 ml/min, column temperature was maintained at 28°C and effluents were monitored at 225 nm. The injection volume was 10 μl.
Results: The retention time of andrographolide and aloe-emodin was found to be 4.57±0.2 min and 12.29±0.2 min, respectively. The markers were resolved using linear responses that were obtained in concentration ranges of 0.5–60 μg/ml with correlation coefficient (r2) of 0.9992 and 0.999 for andrographolide and aloe-emodin, respectively. The precision results were found to be satisfactory, which indicates that the method is precise. The recovery values lie in the range of 98–120% indicating the accuracy of the method.
Conclusion: A novel, simple, accurate, precise, and robust reverse-phase high-performance liquid chromatography method was developed for the simultaneous estimation of andrographolide and aloe-emodin. The developed method can be used for analysis of the formulations containing these phytoconstituents.
2. Shaikh H, Jain V. A novel, simple, rapid RP-HPLC method for simultaneous estimation of ferulic acid, quercetin, piperine and thymol in Ayuvedic formulation. Int J Appl Pharm 2018;10:303-8.
3. EMEA. Quality of Herbal Medicinal Products. Guidelines. European Agency for the Evaluation of Medicinal Products (EMEA): London; 1998.
4. Ong ES. Extraction methods and chemical standardization of botanicals and herbal preparations. J Chromatogr B Analyt Technol Biomed Life Sci 2004;812:23-33.
5. Lokhande S, Chougule A, Patil S, Patil V. Need of herbal drug standardization. Int Ayurvedic Med J 2015;3:874-7.
6. Kotagiri R, Kanaujia A, Singh P, Thakur D. Validated RP-HPLC method for the quantification of andrographolide in toxiroak premix, a polyherbal mycotoxin inhibitor. Int J Pharm Sci Res 2013;4:2623-8.
7. Xu W, Yang W. Assay of andrographolide bulk drug by HPLC. Zhongguo Zhong Yao Za Zhi 2010;35:2113-5.
8. Bhope SG, Kuber VV, Nagore DH, Gaikwad PS, Patil MJ. Development and validation of RP-HPLC method for simultaneous analysis of andrographolide, phyllanthin and hypophyllanthin from herbal hepatoprotective formulation. Acta Chromatogr 2013;25:159-69.
9. Kumaran KS, Thirugnanasambantham P, Vishwanathan S, murthy MS. An HPLC method for the estimation of andrographolide in rabbit serum. Indian J Pharmacol 2003;35:109-12.
10. Mandrioli R, Mercolini L, Ferranti A, Fanali S, Raggi MA. Determination of aloe emodin in Aloe vera extracts and commercial formulations by HPLC with tandem UV absorption and fluorescence detection. Food Chem 2011;126:387-93.
11. Tabin S, Gupta RC, Bansal G, Kamili AN. Comparative HPLC analysis of emodin, aloe emodin and rhein in Rheum emodi of wild and in vitro raised plants. J Pharmacogn Phytochem 2016;5:121-30.
12. Zaffaroni M, Mucignat C, Pecera T, Zagotto G, Frapolli R, D’Iucalci M, et al. High-performance liquid chromatographic assay for the determination of aloe emodin in mouse plasma. J Chromatogr B Analyt Technol Biomed Life Sci 2003;796:113-9.
13. New Livfit Tablet Uses, Side effects, Reviews, and Precautions Alembic Pharma Tablet Wise India. Available from: https://www.tabletwise. com/new-livfit-tablet. [Last accessed on 2019 Sep 18].
14. Joseph J, Solomon J. Andrographis paniculata: A review of its traditional uses, phytochemistry and pharmacology. Med Aromat Plants 2014;3:2-14.
15. Handa SS, Sharma A. Hepatoprotective activity of andrographolide against galactosamine and paracetamol intoxication in rats. Indian J Med Res 1990;92:284-92.
16. Malik MA, Bhat SA, Bilquees F, Sheikh BA, Sidiqui S, Shrivastava P. Rheum emodi as valuable medicinal plant. Int J Gen Med Pharm 2016;5:35-44.
17. Sanders B, Ray AM, Goldberg S, Clark T, Clark T, McDaniel HR, et al. Anti-cancer effects of aloe-emodin: A systematic review. J Clin Transl Res 2017;3:283-96.
18. ICH Harmonised, Tripartite Guideline Validation of Analytical Procedures: Text and Methodology Q2(R1), Geneva; 2005. p. 1-13.
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