Cover Image

Development and Validation of HPLC-PDA Assay method of Frangula emodin

Deborah Duca, Claude Farrugia


Frangula emodin, (1,3,8-trihydroxy-6-methyl-anthraquinone), is one of the anthraquinone derivatives found abundantly in the roots and bark of a number of plant families traditionally used to treat constipation and haemorrhoids. The present study describes the development and subsequent validation of a specific Assay HPLC method for emodin. The separation was achieved on a Waters Symmetry C18, 4.6 × 250 mm, 5 μm particle size, column at a temperature of 35 °C, with UV detection at 287 and 436 nm. An isocratic elution mode consisting of 0.1% formic acid and 0.01% trifluoroacetic acid as the aqueous mobile phase, and methanol was used. The method was successfully and statistically validated for linearity, range, precision, accuracy, specificity and solution stability.

Full Text:



- European Council Directive 2004/24 EC of the European parliament and of the Council of 31 March 2004, amending as regards traditional herbal medicinal products, Directive 2001/83/EC on the Community code relating to medicinal products for human use.

- S. Tseng, H. Lee, L. Chen, C. Wu, C. Wang, Effects of three purgative decoctions on inflammatory mediators. J Ethnopharmacol., 2006,118-24.

- I. Izhaki, Emodin – a secondary metabolite with multiple ecological functions in higher plants. New Phytologist, 2002, 205-217.

- Y. Lee, O. Kang, J. Choi, Y. Oh, J. Keum, S. Kim, Synergistic effect of emodin in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus. Pharm Biol, 2010,1285-1290.

- Y. Liu, N. Shen, C. Liu, Y. Lv, Immunosuppressive effects of emodin: an in vivo and in vitro study. Transplant Proc., 2009, 1837-9.

- J. Xue, W. Ding, Y. Liu, Anti-diabetic effects of emodin involved in the activation of PPARgamma on high-fat diet-fed and low dose of streptozotocin-induced diabetic mice. Fitoterapia, 2010, 173-7.

- W. Lee, S. Ku, D. Lee, T. Lee, B. JS, Emodin-6-O-β-D--glucoside Inhibits High-Glucose-Induced Vascular Inflammation. Inflammation. 2014, 37(2):306-13

- G. Meng, Y. Liu, C. Lou, H. Yang, Emodin suppresses lipopolysaccharide-induced pro-inflammatory responses and NF-kappaB activation by disrupting lipid rafts in CD14-negative endothelial cells. Br J Pharmacol, 2010, 1628-1644.

- J. Lai, J. Chang, C. Wen, Emodin induces a reactive oxygen species-dependent and ATM-p53-Bax mediated cytotoxicity in lung cancer cells. Eur J Pharmacol, 2009, 1-9.

- C. Hsu, Y. Hsu, Y. Tsai, Emodin inhibits the growth of hepatoma cells: finding the common anti-cancer pathway using Huh7, Hep3B, and HepG2 cells. Biochem Biophys Res Commun, 2010, 473-478.

- J. Cai, X. Niu, Y. Chen, Q. Hu, G. Shi, H. Wu, J. Wang, J. Yi, Emodin-induced generation of reactive oxygen species inhibits RhoA activation to sensitize gastric carcinoma cells to anoikis. Neoplasia, 2008, 41-45.

- R. Räisänen, H. Björk, P. Hynninen, Two-dimensional TLC separation and mass spectrometric identification of anthraquinones isolated from the fungus Dermocybe sanguinea. Z Naturforsch C, 2000, 55(3-4), 195-202.

- B. Jiang, X. Gao, K. Bi, XH. Chen, Simultaneous determination of anthraquinones in Liuwei'anxiao capsules by HPLC. Zhong Yao Cai., 2009, 32(6), 975-7.

- Y. Wang, X. Huang, Q. Liang, R. Fan, F. Qin, J. Luo, L. Gao, Simultaneous quantitation of seven anthraquinones in aqueous extract of rhubarb and analysis for absorption in cerebrospinal fluid of patients with traumatic brain injury using UPLC-PDA method. Journal of Medicinal Plants, 2011, 2879-2885.

- D. Yan, Y. Ma, Simultaneous quantification of five anthraquinones in rat plasma by high-performance liquid chromatography with fluorescence detection. Biomed Chromatogr., 2007, 21(5), 502-7.

- S. Mueller, M. Schmitt, W. Dekant, H. Stopper, J. Schlatter, P. Schreier, W. Lutz, Occurrence of emodin, chrysophanol and physcion in vegetables, herbs and liquors. Genotoxicity and anti-genotoxicity of the anthraquinones and of the whole plants. Food Chem Toxicol., 1999, 37(5), 481-91.

- L.-X. Peng, J.B. Wang, L.X. Hu, Rapid and simple method for the determination of emodin in tartary buckwheat (Fagopyrum tataricum) by high performance liquid chromatography coupled to a diode array detector. Journal of Agricultural and Food Chemistry, 2013, Issue 61, p. 854−857.

- H.M. Lu, Y. Liang, W. Ni, R. Man, Optimization of extraction and determination of emodin from Polygonum cuspidatum Sieb. et Zucc. products by HPLC-DAD. J. Cent. South Univ. Technol., 2006, 658-662.

- International Conference on Harmonization (ICH) guidelines ICH Q2(R1), Validation of analytical procedures: text and methodology, 2005.

- J. Mehta, Separation and characterization of anthraquinone derivatives from Cassia fistula using chromatographic and spectral techniques. Int. J. Chem. Sci., 2002, 306-316.

- S. Genovese, F. Tammaro, L. Menghini, G. Carlucci, F. Epifanoa and M. Locatellia, Comparison of three different extraction methods and HPLC determination of the anthraquinones aloe-emodine, emodine, rheine, chrysophanol and physcione in the bark of Rhamnus alpinus L. Rhamnaceae. Phytochem. Anal., 2010, 261–267

- Lu et al., Preparation of a nano emodin transfersome and study on its anti-obesity mechanism in adipose tissue of diet-induced obese rats, Journal of Translational Medicine, 2014, 12:72

- J. Wang, H. Li, C. Jin, Y. Qu, X. Xiao, Development and validation of a UPLC method

for quality control of rhubarb-based medicine: Fast simultaneous determination of five anthraquinone derivatives. Journal of Pharmaceutical and Biomedical Analysis, 2008, 765-770.

- Y-B Shi, Y-P Shi, Y-B Yang, G. Feng, Simultaneous determination of tetrehydropalmatine, magnolol, emodin and chrysophanol in Chinese Herbal Preparation by RP-HPLC-PDA. Chromatographia., 2007, 601-606.

- R. Song, F. Xu, Z. Zhang, Y. Liu, H. Dong, Y. Tian, Structural elucidation of in vitro metabolites of emodin by liquid chromatography–tandem mass spectrometry. Biomedical Chromatography, 2008, 1230-1236.

- D. He, B. Chen, Q. Tian, S. Yao, Simultaneous determination of five anthraquinones in medicinal plants and pharmaceutical preparations by HPLC with fluorescence detection. Journal of Pharmaceutical and Biomedical Analysis, 2009, 1123-1127.

- D. Wang, G. Yang, X. Song, Determination of pKa values of anthraquinone compounds by capillary electrophoresis. Electrophoresis, 2001, 464-469.

- Z. Marković, N. Manojlović, S. Zlatanović, Electronic Absorption Spectra of Substituted Anthraquinones and Their Simulation Using ZINDO/S Method. Journal of the Serbian Society for Computational Mechanics, 2008, 73-79.

- X. Chu, A. Sun, R. Liu, Preparative isolation and purification of five compounds from the Chinese medicinal herb Polygonum cuspidatum Sieb. et Zucc by high-speed counter-current chromatography. J Chromatogr A., 2005, 1097(1-2):33-9.

–S. Guo, B. Feng, R. Zhu, J. Ma, Wang W., Preparative isolation of three anthraquinones from Rumex japonicus by High-Speed Counter-Current Chromatography. Molecules, 2011, 16, 1201-1210


Copyright (c) 2016 Mediterranean Journal of Chemistry