STABILITY-INDICATING REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF APIGENIN AND LUTEOLIN FROM ACHILLEA MILLEFOLIUM LINN
Objective: A stability-indicating reverse-phase high-performance liquid chromatographic method was developed and validated for the analysis of apigenin and luteolin. The degradation behavior of apigenin and luteolin was investigated under different stress conditions as recommended by the International Conference on Harmonization (ICH).
Methods: In the present study, a reversed-phase high-performance liquid chromatography method was developed and the resolution of the plant constituents was successfully achieved using Hibar Lichrospher C8 column with ultraviolet detector at a wavelength of 269 nm. The mobile phase consisted of methanol and 0.5% trifluoroacetic acid (80:20 v/v) at a flow rate of 1.0 ml/min. Both apigenin and luteolin were subjected to various stress degradation studies such as oxidation, acid and alkaline hydrolysis, and photolytic degradation.
Results: The proposed method was found to be linear (1–5 μg/ml) with the linear correlation coefficient of R2=0.99. Although the degradation products of stressed conditions were not identified, the methods were able to detect the changes due to stress condition.
Conclusion: The method provides good sensitivity and excellent precision and reproducibility. Forced degradation studies on apigenin and luteolin give information about their storage and intrinsic stability conditions considering the advanced pharmaceutical aspects of formulations.
2. Sachan AK, Kumar A. Stability testing of herbal products-A review. J Chem Pharm Res 2015;7:511-4.
3. Ahuja S, Alsante KM. Handbook of Isolation and Characterisation of Impurities in Pharmaceuticals. San Diego USA: Academic Press; 2003.
4. Naidu KR, Kale UN, Shingare MS. Stability indicating RP-HPLC method for simultaneous determination of amlodipine and benazepril hydrochloride from their combination drug product. J Pharm Biomed Anal 2005;39:147-55.
5. Blessy M, Patel RD, Prajapati PN, Agrawal YK. Development of forced degradation and stability indicating studies of drugs-A review. J Pharm Anal 2014;4:159-65.
6. International Federation of Pharmaceutical Manufacturers & Associations. ICH, Q1A Stability Testing of New Drug Substances and Products: Proceedings of the International Conference on Harmonization. Geneva: International Federation of Pharmaceutical Manufacturers & Associations; 2000.
7. Bimbiraite K, Ragazinskiene O, Maruska A, Kornysova O. Comparison of the chemical composition of four yarrow (Achillea millefolium L.) morphotypes. Biologija 2008;54:208-12.
8. Raote A, Jangid A, Tale R, Vaidya V. Liquid chromatography-Tandem mass spectrometric method for simultaneous determination of rutin and quercetin from leaves of Artocarpus lakoocha Roxb. Int J Pharm Bio Sci 2011;2:848-53.
9. Devaraj VC, Krishna BG, Viswanatha GL. Simultaneous determination of quercetin, rutin and kaempferol in the leaf extracts of Moringa oleifera lam. And Raphinus sativus linn. By liquid chromatography-tandem mass spectrometry. Zhong Xi Yi Jie He Xue Bao 2011;9:1022-30.
10. Ashok Kumar BS, Lakshman K, Jayaveera KN, Vamshi Krishna N, Mnjunath M, Suresh MV. Estimation of rutin and quercetin in Amaranthus viridis Linn by HPLC. Asian J Exp Sci 2009;23:51-4.
11. Shen H, Guo Q, Fang H, Wang Y, Jin M. Determination of quercetin, luteolin, apigenin and acacetin in flos Chrysanthemi indici by RP-HPLC. Zhongguo Zhong Yao Za Zhi 2010;35:191-3.
12. Hamrapurkar P, Patil P, Desai M, Phale M, Pawar S. Stress degradation studies and development of a validated stability-indicating-assay-method for determination of diacerein in presence of degradation products. Pharm Methods 2011;2:30-5.
13. Sarin YK. Problems in standardization of botanicals used by indigenous drug industry in India. Indian J Nat Prod 1993;9:12-7.
14. Rajendra BP, Tushar AD, Vijay RP. Stability indicating HPLC method for dapoxetine HCl in bulk and in formulation. Int J Pharm Pharm Sci 2014;6:687-90.
15. Kuna M, Dannana GS. Development and validation of stability indicating reverse-phase high performance liquid chromatography method for the simultaneous quantification of saquinavir, ritonavir and amprenavir. Asian J Pharm Clin Res 2018;11:390-6.
16. Alsante KM, Ando A, Brown R, Ensing J, Hatajik TD, Kong W, et al. The role of degradant profiling in active pharmaceutical ingredients and drug products. Adv Drug Deliv Rev 2007;59:29-37.
17. Reynolds DW, Facchine KL, Mullaney JF, Alsante KM, Hatajik TD, Motto MG. Available guidance and best practices for conducting forced degradation studies. Pharm Technol 2002;26:48-56.
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.