Abstract
A method for rapid preparative isolation and screening of antioxidants has been developed by combining preparative HSCCC with on-line radical scavenging detection by use of DPPH· (1,1-diphenyl-2-picrylhydrazyl) as model radical. The HSCCC effluent was split into two parts. The major part was collected and the rest was introduced, through a switch valve, directly into a reaction coil for on-line evaluation of antioxidant activity. Each of the antioxidants separated by HSCCC reacted postcolumn with the DPPH· solution in methanol and was observed as a negative peak at 517 nm of size corresponding to its antioxidant activity. The on-line method was successfully used for preparative isolation and screening of antioxidant components from an ethyl acetate extract of the traditional Chinese medicine Selaginella moellendorffii. A total of 114.7 mg amentoflavone, 12.4 mg hinokiflavone, 7.2 mg podocarpusflavone A, and 1.1 mg ginkgetin, identified as the main radical scavengers, were obtained with purity of 99.4, 98.6, 98.9, and 99.2%, respectively, as determined by LC, from 1.0 g ethyl acetate extract. Their chemical structures were identified by MS and by UV and NMR spectroscopy.
Similar content being viewed by others
References
Gilgun-Sherki Y, Rosenbaum Z, Melamed E, Offen D (2002) Pharmacol Rev 54:271–284. doi:10.1124/pr.54.2.271
Chang YC, Lee FW, Chen CS, Huang ST, Tsai SH, Huang SH et al (2007) Free Radic Biol Med 43:1541–1551. doi:10.1016/j.freeradbiomed.2007.08.018
Choi HS, Song HS, Ukeda H, Sawamura M (2000) J Agric Food Chem 48:4156–4161. doi:10.1021/jf000227d
Siddhuraju P (2007) LWT 40:982–990. doi:10.1016/j.lwt.2006.07.010
Shahidi F, Alasavar C, Liyana-Pathirana CM (2007) J Agric Food Chem 55:1212–1220. doi:10.1021/jf062472o
Shyamala BN, Naidu MM, Sulochanamma G, Srinivas P (2007) J Agric Food Chem 55:7738–7743. doi:10.1021/jf071349±
Souza JNS, Silva EM, Loir A, Rees J, Rogez H, Larondella Y (2008) Food Chem 106:331–339. doi:10.1016/j.foodchem.2007.05.011
Jiangsu New Medical College (1988) Dictionary of Chinese Materia Medica (Zhongyao Dacidian). Shanghai Scientific and Technological Publishers, Shanghai, pp 818–819
Benedek B, Weniger B, Parejo I, Bastida J, Arango GJ, Lobstein A et al (2006) Arzneim Forsch 56:661–664
Mukherjee PK, Ahamed KFHN, Kumar V, Mukherjee K, Houghton PJ (2007) Behav Brain Res 178:221–228. doi:10.1016/j.bbr.2006.12.025
Sun CM, Syu WJ, Huang YT, Chen CC, Ou JC (1997) J Nat Prod 60:382–384. doi:10.1021/np960608e
Mbukwa E, Chacha M, Majinda RRT (2007) Arkivoc 104–112.
Chen DZ, Yu JG (1986) Zhongcaoyao 17:4
Ito Y (1981) J Chromatogr A 214:122–125. doi:10.1016/S0021-9673(00)80907-3
Ito Y (1991) J Chromatogr A 538:3–25. doi:10.1016/S0021-9673(01)91617-6
Okunji C, Komarnytsky S, Fear G, Poulev A, Ribnicky DM, Awachie PI et al (2007) J Chromatogr A 1151:45–50. doi:10.1016/j.chroma.2007.02.085
Gu DY, Yang Y, Zhong J, Asia HA, Zhang TY (2007) Chromatographia 66:949–951. doi:10.1365/s10337-007-0432-2
Gutzeit D, Wray V, Winterhalter P, Jerz G (2007) Chromatographia 65:1–7. doi:10.1365/s10337-006-0105-6
Pukalskas A, van Beek TA, De Waard P (2005) J Chromatogr A 1074:81–88. doi:10.1016/j.chroma.2005.03.089
Kosar M, Dorman HJD, Baser KHC, Hiltunen R (2004) J Agric Food Chem 52:5004–5010. doi:10.1021/jf0496189
Koleva II, Niderländer HAG, van Beek TA (2000) Anal Chem 72:2323–2328. doi:10.1021/ac9912451
Pérez-Bonilla M, Salido S, van Beek TA, Linares-Palomino PJ, Altarejos J, Nogueras M et al (2006) J Chromatogr A 1112:311–318. doi:10.1016/j.chroma.2005.12.055
Bandonienè D, Murkovic M (2002) J Agric Food Chem 50:2482–2487. doi:10.1021/jf011475s
Ruzicka J, Hansen EH (1978) Anal Chim Acta 99:37–76. doi:10.1016/S0003-2670(01)84498-6
Chen LJ, Games DE, Jones J (2003) J Chromatogr A 988:95–105. doi:10.1016/S0021-9673(02)01954-4
Miki K, Nagai T, Suzuki K, Tsujimura R, Koyama K, Kinoshita K et al (2007) Bioorg Med Chem Lett 17:772–775. doi:10.1016/j.bmcl.2006.10.075
Weniger B, Vonthron-Sénécheau C, Kaiser M, Brun R, Anton R (2006) Phytomedicine 13:176–180. doi:10.1016/j.phymed.2004.10.008
Carbonezi CA, Hamerski L, Gunatilaka AAL, Cavalheiro A, Castro-Gamboa L, Silva DHS et al (2007) Rev Bras Farmacogn 17:319–324. doi:10.1590/S0102-695X2007000300003
Joyeux M, Lobstein A, Anton R, Mortier F (1995) Planta Med 61:126–129. doi:10.1055/s-2006-958030
Markham KR, Sheppard C, Geiger H (1987) Phytochemistry 26:3335–3337. doi:10.1016/S0031-9422(00)82499-1
Acknowledgments
The authors wish to acknowledge the assistance of Professor Qizhen Du for his excellent technical support. This work was supported by the Postdoctoral Science Foundation of Central South University.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Shi, S., Zhou, H., Zhang, Y. et al. Hyphenated HSCCC–DPPH· for Rapid Preparative Isolation and Screening of Antioxidants from Selaginella moellendorffii . Chroma 68, 173–178 (2008). https://doi.org/10.1365/s10337-008-0716-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1365/s10337-008-0716-1