Abstract
A method for the determination of artepillin-C using HPLC coupled with photodiode array (PDA) detector was developed. Artepillin-C is abundantly present in Brazilian propolis of Baccharis species origin. The chromatographic separation was performed on a Mightysil RP-18 GP II column (150 mm × 4.6 mm, 5.0 ?m; Kanto Chemical Company Limited), and column temperature was maintained at 40°C. The mobile phase was a linear gradient elution of 0.5% aqueous acetic acid (A) and acetonitrile (B) at a flow rate of 1.0 ml/min. The chromatogram was monitored at 320 nm. The calibration range of artepillin-C was 0.75-2500 ?g/ml (r2 = 0.9991). The limit of detection (LOD) and limit of quantification (LOQ) were 0.50 ?g/ml and 0.75 ?g/ml, respectively. The intra-day and inter-day precision of the assay (RSD) were in the range 1.28-5.60% and 1.45-6.75%, respectively. The artepillin-C content in Brazilian propolis is an important factor in ensuring the quality of Brazilian propolisof Baccharis species origin. This method was used to determine artepillin-C content in Brazilian propolis.
References
Bankova VS, De Castro SL, Marcucci MC. Propolis: recent advances in chemistry and plant origin. Apidologie 2000; 31: 3-15. http://dx.doi.org/10.1051/apido:2000102
Scazzocchio F, D’Auria FD, Alessandrini D, Pantanella F. Multifactorial aspects of antimicrobial activity of propolis. Microbiol Res 2006; 161: 327-33. http://dx.doi.org/10.1016/j.micres.2005.12.003
Shimizu T, Hino A, Tsutsumi A, Park YK, Watanabe W, Kurokawa M. Anti-influenza virus activity of propolis in vitro and its efficacy against influenza infection in mice. Antivir Chem Chemother 2008; 19(1): 7-13.
Tan-no K, Nakajima T, Shoji T, et al. Anti-inflammatory effect of propolis through inhibition of nitric oxide production on carrageenin-induced mouse paw edema. Biol Pharm Bull 2006; 29(1): 96-99. http://dx.doi.org/10.1248/bpb.29.96
Murad JM, Calvi SA, Soares AMVC, Bankova V, Sforcin JM. Effects of propolis from Brazil and Bulgaria on fungicidal activity of macrophages against Paracoccidioides brasiliensis. J Ethnopharmacol 2002; 79: 331-34. http://dx.doi.org/10.1016/S0378-8741(01)00404-4
Shimizu K, Ashida H, Matsuura Y, Kanazawa K. Antioxidant bioavailability of artepillin-C in Brazilian propolis. Arch Biochem Biophys 2004; 424: 181-88. http://dx.doi.org/10.1016/j.abb.2004.02.021
Nakajima Y, Shimazawa M, Mishima S, Hara H. Water extract of propolis and its main constituents, caffeoylquinic acid derivatives, exert neuroprotective effects via antioxidant actions. Life Sci 2007; 80: 370-77. http://dx.doi.org/10.1016/j.lfs.2006.09.017
Izuta H, Narahara Y, Shimazawa M, Mishima S, Kondo S, Hara H. 1,1-Diphenyl-2-picrylhydrazyl radical scavenging activity of bee products and their constituents determined by ESR. Biol Pharm Bull 2009; 32(12): 1947-51. http://dx.doi.org/10.1248/bpb.32.1947
Moreira L, Dias LG, Pereira JA, Estevinho L. Antioxidant properties, total phenols and pollen analysis of propolis samples from Portugal. Food Chem Toxicol 2008; 46(11): 3482-85. http://dx.doi.org/10.1016/j.fct.2008.08.025
Yoshikawa T, Tsuji T. The doctor’s complete guide to alternative medicines. Tokyo, Kodansha 2004; pp. 114-115.
Nakamura J, Matsuka M. Propolis as a material for complementary and alternative medicine. Jpn J Complement Alternat Med 2005; 2: 45-57. http://dx.doi.org/10.1625/jcam.2.45
Furukawa S, Takagi N, Ikeda T, et al. Two novel long-chain alkanoic acid esters of lupeol from Alecrim-propolis. Chem Pharm Bull 2002; 50(3): 439-40. http://dx.doi.org/10.1248/cpb.50.439
Funari CSD, Ferro VDO, Mathor MB. Analysis of propolis from Baccharis dracunculifolia DC. (Compositae) and its effects on mouse fibroblasts. J Ethnopharmacol 2007; 111(2): 206-12. http://dx.doi.org/10.1016/j.jep.2006.11.032
Kumazawa S, Yoneda M, Shibata I, Kanaeda J, Hamasaka Y, Nakayama T. Direct evidence for the plant origin of Brazilian propolis by the observation of honeybee behavior and phytochemical analysis. Chem Pharm Bull 2003; 51(6): 740-42. http://dx.doi.org/10.1248/cpb.51.740
Estrada GOD, Mendes da Silva JF, Antunes OAC. Artepillin C: A Review. Lett Drug Des Discov 2008; 5: 88-92. http://dx.doi.org/10.2174/157018008783928436
Tazawa S. Analysis of the constituents, chemical evaluation and tyrosinase inhibition of propolis. Fragrance J 2002; 3: 25-32.
Nafady AM, El-Shanawany MA, Mohamed MH, et al. Cyclodextrin-enclosed substances of Brazilian propolis. Chem Pharm Bull 2003; 51(8): 984-85. http://dx.doi.org/10.1248/cpb.51.984
Gardana C, Scaglianti M, Pietta P, Simonetti P. Analysis of the polyphenolic fraction of propolis from different sources by liquid chromatography-tandem mass spectrometry. J Pharm Biomed Anal 2007; 45(3): 390-99. http://dx.doi.org/10.1016/j.jpba.2007.06.022
Kumazawa S, Yoneda M, Nakayama T. Constituents in Brazilian propolis and its plant of origin. Foods and Food Ingredients J Jpn 2004; 209(2): 132-40.
Han L, Liu K, Wang S, Wang X, Hou H. Determination of artepillin-C in propolis by HPLC. Zhongguo Yaoshi 2008; 22(4): 312-14.
Matsuda AH, Bicudo De Almeida-Muradian L. Validated method for the quantification of artepillin-C in Brazilian propolis. Phytochem Anal 2008; 19(2): 179-83. http://dx.doi.org/10.1002/pca.1043
![Creative Commons License](http://i.creativecommons.org/l/by-nc/4.0/88x31.png)
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Copyright (c) 2012 Yasuhito Nobushi, Naoki Oikawa, Yuzo Okazaki, Shigetoshi Tsutsumi, Yong Kun Park, Masahiko Kurokawa , Ken Yasukawa